Methods, compounds, and compositions for reducing body fat and modulating fatty acid metabolism

ABSTRACT

Methods, pharmaceutical compositions, and compounds for reducing body weight, modulating body lipid metabolism, and reducing food intake in mammals are provided. The compounds of the invention include fatty acid ethanolamide compounds, homologues and analogs of which the prototype is the endogenous fatty acid ethanolamide, oleoylethanolamide.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority of U.S. Patent Application No.60/336,289 filed Oct. 31, 2001 and U.S. Patent Application No.60/279,542 filed Mar. 27, 2001. The contents of which are eachincorporated herein by reference.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

[0002] This invention was made with government support under Grant No.DA 12653, awarded by the National Institute of Health. The Governmenthas certain rights in this invention.

FIELD OF THE INVENTION

[0003] This invention relates to fatty acid ethanolamides, theirhomologues, and their analogs and to their use as pharmacologicallyactive agents to reduce body fat, reduce food consumption, and modulatelipid metabolism.

BACKGROUND OF THE INVENTION

[0004] Obesity is a worldwide health challenge occuring at alarminglevels in the United States and other developed nations. About 97million adults in the United States are overweight. Of these 40 millionare obese. Obesity and overweight greatly increase the risk of manydiseases. Hypertension; type 2 diabetes; dyslipidemia; coronary heartdisease; stroke; gallbladder disease; osteoarthritis; sleep apnea andother respiratory problems; and endometrial, breast, prostate, and coloncancers have been associated with higher body weights. Persons withhigher body weights also suffer from a higher all-cause death rate.According to the National Institutes of Health about 280,000 adultdeaths in the United States each year may be attributed in part toobesity.

[0005] Weight loss is desirable in the case of obesity and overweightindividuals. Weight loss can help to prevent many of these harmfulconsequences, particularly with respect to diabetes and cardiovasculardisease (CVD). Weight loss may also reduce blood pressure in bothoverweight hypertensive and non-hypertensive individuals; serumtriglycerides levels and increases the beneficial high-densitylipoprotein (HDL)-form of cholesterol. Weight loss also generallyreduces somewhat the total serum cholesterol and low-density lipoprotein(LDL)-cholesterol levels. Weight loss may also reduce blood glucoselevels in overweight and obese persons.

[0006] While weight loss is desirable, it is hard to achieve. Manytreatments for the management of overweight and obesity and themaintenance of weight loss exist. However, recidivism is rampant.Approximately, 40 percent of women and 24 percent of men are trying toactively lose weight at any given time. These treatments includelow-calorie diets and low-fat diets; increased physical exercise;behavioral therapies directed toward reducing food intake,pharmacotherapy; surgery; and combinations of the above.

[0007] The pharmacopoea of weight loss is relatively bare. Drugs such assibutramine, dexfenfluramine, orlistat, phenylpropanolamine,phenteramine, or fenfluramine can facilitate weight loss in obese adultswhen used for prolonged periods. In general, however, the safety oflong-term administration of pharmaco-therapeutic weight loss agents isunknown. For instance, recently due to concerns about valvular heartdisease observed in patients, fenfluramine and dexfenfluramine have beenwithdrawn from the market. In the face of the slim pharmacopoea and thehigh prevalence of obesity and overweight, there is a need for newpharmaceutical methods and compositions to promote and maintain weightloss.

[0008] Fatty acid ethanolamides (FAE) are unusual components of animaland plant lipids, and their concentrations in non-stimulated cells aregenerally low (Bachur et al., J. Biol. Chem., 240:1019-1024 (1965);Schmid et al., Chem. Phys. Lipids, 80:133-142 (1996); Chapman, K. D.,Chem. Phys. Lipids, 108:221-229 (2000)). FAE biosynthesis can be rapidlyenhanced, however, in response to a wide variety of physiological andpathological stimuli, including exposure to fungal pathogens in tobaccocells (Chapman et al., Plant Physiol., 116:1163-1168 (1998)), activationof neurotransmitter receptors in rat brain neurons (Di Marzo et al.,Nature, 372:686-691 (1994); Giuffrida et al., Nat. Neurosci., 2:358-363(1999)) and exposure to metabolic stressors in mouse epidermal cells(Berdyshev et al., Biochem. J., 346:369-374 (2000)). The mechanismunderlying stimulus-dependent FAE generation in mammalian tissues isthought to involve two concerted biochemical reactions: cleavage of themembrane phospholipid, N-acyl phosphatidylethanolamine (NAPE), catalyzedby an unknown phospholipase D; and NAPE synthesis, catalyzed by acalcium ion- and cyclic AMP-regulated N-acyltransferase (NAT) activity(Di Marzo et al., Nature, 372:686-691 (1994); Cadas et al., J.NeuroSci., 6:3934-3942 (1996); Cadas et al., H., J. Neurosci.,17:1226-1242, (1997)).

[0009] The fact that both plant and animal cells release FAEs in astimulus-dependent manner suggests that these compounds may playimportant roles in cell-to-cell communication. Further support for thisidea comes from the discovery that the polyunsaturated FAE, anandamide(arachidonylethanolamide), is an endogenous ligand for cannabinoidreceptors (Devane et al., Science, 258:1946-1949 (1992))—Gprotein-coupled receptors expressed in neurons and immune cells, whichrecognize the marijuana constituent Δ⁹-tetrahydrocannabinol (Δ⁹-THC)(for review, see reference (Pertwee, R. G., Exp. Opin. Invest. Drugs,9:1553-1571 (2000)).

[0010] Two observations make it unlikely that other FAEs alsoparticipate in cannabinoid neurotransmission. The FAE family iscomprised for the most part of saturated and monounsaturated species,such as palmitylethanolamide and oleoylethanolamide, which do notsignificantly interact with cannabinoid receptors (Devane et al.,Science, 258:1946-1949 (1992); Griffin et al., J. Pharmacol. Exp. Ther.,292:886-894. (2000)). Second, when the pharmacological properties of theFAEs have been investigated in some detail, as is the case withpalmitylethanolamide, such properties have been found to differ fromthose of Δ⁹-THC and to be independent of activation of known cannabinoidreceptor subtypes (Calignano et al., Nature, 394:277-281 (1998)). Thus,the biological significance of the FAEs remains elusive.

[0011] Oleoylethanolamide (OEA) is a natural analogue of the endogenouscannabinoid anandamide. Like anandamide, OEA is produced in cells in astimulus-dependent manner and is rapidly eliminated by enzymatichydrolysis, suggesting a role in cellular signaling. However, unlikeanandamide, OEA does not activate cannabinoid receptors and itsbiological functions were here-to-fore essentially unknown.

[0012] There is a need for additional methods and agents to treatobesity and overweight as well as to maintain weight loss. The presentinvention meets this need by providing novel methods and pharmaceuticalcompositions related to our instant discovery that oleoylethanolamide(OEA) and other fatty acid ethanolamide compounds (e.g.,palmitylethanolamide, elaidylethanolamide)) can reduce appetite, foodintake, body weight, and body fat and alter fat metabolism.

SUMMARY OF THE INVENTION

[0013] The present invention provides compounds, compositions, andmethods for reducing body fat and for treating or preventing obesity,and overweight in mammals and the diseases associated with these healthconditions. In one aspect, the invention provides methods for reducingbody fat or body weight and for treating or preventing obesity oroverweight and for reducing food intake by administration ofpharmaceutical compositions comprising a fatty acid alkanolamidecompound, homologue, or analog in an amount sufficient to reduce bodyfat, body weight or prevent body fat or body weight gain. In otheraspects, the invention is drawn to the fatty acid ethanolamidecompounds, homologues, analogs; and their pharmaceutical compositionsand such methods of use.

[0014] In other embodiments, the fatty acid moiety of the fatty acidalkanolamide or ethanolamide compound, homologue, or analog may besaturated or unsaturated, and if unsaturated may be monounsaturated orpolyunsaturated.

[0015] In some embodiments, the fatty acid moiety of the fatty acidalkanolamide compound, homologue, or analog is a fatty acid selectedfrom the group consisting of oleic acid, palmitic acid, elaidic acid,palmitoleic acid, linoleic acid, alpha-linolenic acid, andgamma-linolenic acid. In certain embodiments, the fatty acid moietieshave from twelve to 20 carbon atoms.

[0016] Other embodiments are provided by varying the hydroxyalkylamidemoiety of the fatty acid amide compound, homologue or analog. Theseembodiments include the introduction of a substituted or unsubstitutedlower (C₁-C₃) alkyl group on the hydroxyl group of an alkanolamide orethanolamide moiety so as to form the corresponding lower alkyl ether.In another embodiment, the hydroxy group of the alknaolamide orethanolamide moiety is bound to a carboxylate group of a C₂ to C₆substituted or unsubstituted alkyl carboxylic acid to form thecorresponding ester of the fatty acid ethanolamide. Such embodimentsinclude fatty acid alkanolamide and fatty acid ethanolamides in esterlinkage to organic carboxylic acids such as acetic acid, propionic acid,and butanoic acid. In one embodiment, the fatty acid alkanolamide isoleoylalkanolamide. In a further embodiment, the fatty acid alkanolamideis oleoylethanolamide.

[0017] In still another embodiment, the fatty acid ethanolamidecompound, homologue, or analog further comprises a substituted orunsubstituted lower alkyl (C₁-C₃) group covalently bound to the nitrogenatom of the fatty acid ethanolamide.

[0018] In another aspect, the invention provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient and acompound, or its pharmaceutically acceptable salt, having the formula:

[0019] In this formula, n is from 0 to 5 and the sum of a and b can befrom 0 to 4. Z is a member selected from —C(O)N(R^(o))—; —(R^(o))NC(O)—;—OC(O)—; —(O)CO—; O; NR^(o); and S, in which R¹ and R² are independentlyselected from the group consisting of unsubstituted or unsubstitutedalkyl, hydrogen, substituted or unsubstituted C₁-C₆ alkyl, substitutedor unsubstituted lower (C₁-C₆) acyl, homoalkyl, and aryl. Up to fourhydrogen atoms of either or both the fatty acid portion and ethanolamineportion of the compound may also be substituted by methyl or a doublebond. In addition, the molecular bond between carbons c and d may beunsaturated or saturated. In some embodiments, the fatty acidethanolamide of the above formula is a naturally occurring compound.

[0020] In other aspects of the invention, the methods and compositionsemploy fatty acid ethanolamide and fatty acid alkanolamide compounds,homologs and analogs for reducing body weight in which the compounds,homologs and analogs cause weight loss when administered to test animals(e.g., rats, mice, rabbits, hamsters, guinea pigs).

[0021] In still other aspects, the invention is drawn to methods ofusing arylthiazolidinedione compounds and heteroaryl and aryl oxyaceticacid type compounds to reduce body fat, body weight and appetite.

[0022] Still other aspects of the invention address methods of using andadministering the subject compounds and compositions for reducing bodyweight or reducing appetite or reducing food intake or causinghypophagia in mammals (e.g., humans, cats or dogs). The subjectcompositions may be administered by a variety of routes, includingorally.

BRIEF DESCRIPTION OF THE DRAWINGS

[0023]FIG. 1. Starvation increases circulating oleoylethanolamide levelsin rats: (a) time course of the effects of food deprivation on plasmaoleoylethanolamide (oleylethaolamide, OEA) levels; (b) effect of waterdeprivation (18 h) on plasma oleoylethanolamide levels; (c) effect offood deprivation (18 h) on oleoylethanolamide levels in cerebrospinalfluid (CSF); (d) time course of the effects of food deprivation onplasma anandamide (arachidonylethanolamide, ABA) levels; (e) effect ofwater deprivation (18 h) on anandamide plasma levels; (f) effect of fooddeprivation (18 h) on anandamide levels in CSF. Results are expressed asmean ±s.e.m.; asterisk, P<0.05; two asterisks, P<0.01, n=10 per group.

[0024]FIG. 2. Adipose tissue is a primary source of circulatingoleoylethanolamide: starvation-induced changes in N-acyltransferase(NAT) and fatty acid amide hydrolase (FAAH) activities in various rattissues. (a) fat; (b) brain; (c) liver; (d) stomach; (e) smallintestine. Empty bars, free-feeding animals; filled bars, 18-h fastedanimals. Activities are in pmol/mg protein/min. Asterisk, P<0.05, n=3.

[0025]FIG. 3. Adipose tissue is a primary source of circulatingoleoylethanolamide: starvation-induced changes in NAPE andoleoylethanolamide (oleoylethanolamide, OEA) content in adipose andliver tissues. (a) structures of the oleoylethanolamide precursorsalk-1-palmitoenyl-2-arachidonyl-sn-glycero-phosphoethanolamine-N-oleyl(left panel, NAPE 1) andalk-1-palmityl-2-arachidonyl-sn-glycero-phosphoethanolamine-N-oleyl(right panel, NAPE 2); (b) representative HPLC/MS tracings for selectedions characteristic of NAPE 1 (left panel, m/z=987, deprotonatedmolecule, [M−H]⁻) and NAPE 2 (right panel, m/z=1003, [M−H]⁻) infree-feeding (top) and 18-h fasting rats (bottom); (c) food deprivation(18 h) increases the content of NAPE species in fat and decreases it inliver. All identifiable NAPE species were quantified, including theoleoylethanolamide precursors NAPEl and NAPE 2, and the PEA precursorNAPE 3; (d) food deprivation (18 h) increases oleoylethanolamide contentin fat and liver. Empty bars, free-feeding animals; filled bars, 18-hfasted animals. Asterisk, P<0.05, Student's t test; n=3.

[0026]FIG. 4. Oleoylethanolamide/pranamide selectively suppresses foodintake: (a) dose-dependent effects of oleoylethanolamide(oleoylethanolamide/OEA/pranamide) (i.p., empty squares),elaidylethanolamide (empty circles), PEA (triangles), oleic acid (filledsquares) and anandamide (filled circles) on food intake in 24-hfood-deprived rats. Vehicle alone (70% DMSO in saline, 1 ml per kg,i.p.) had no significant effect on acute food intake; (b) time course ofthe hypophagic effects of oleoylethanolamide (20 mg per kg, i.p.)(squares) or vehicle (lozenges) on food intake. (c) effects of vehicle(V), lithium chloride (LiCl, 0.4 M, 7.5 ml per kg) or oleoylethanolamide(20 mg per kg) in a conditioned taste aversion assay. Empty bars, waterintake; filled bars, saccharin intake. Effects of vehicle (V) oroleoylethanolamide (5 or 20 mg per kg) on: (d) water intake (expressedin ml per 4 h); (e) body temperature; (f) latency to jump in the hotplate analgesia test; (g) percent time spent in open arms in theelevated plus maze anxiety test; (h) number of crossings in the openfield activity test; (i) number of operant responses for food. Asterisk,P<0.05, n=8-12 per group.

[0027]FIG. 5. Effects of subchronic oleoylethanolamide administration onfood intake and body weight: (a) effects of oleoylethanolamide(oleoylethanolamide, OEA) (5 mg per kg, i.p. once a day) (empty bars) orvehicle (5% Tween 80/5% propyleneglycol in sterile saline; filled bars)on cumulative food intake; (b) time course of the effects ofoleoylethanolamide (triangles) or vehicle (squares) on body weightchange; (c) effects of oleoylethanolamide or vehicle on net body weightchange; (d) effects of oleoylethanolamide (5 mg per kg) or vehicle oncumulative water intake. Asterisk, P<0.05; two asterisks, P<0.01, n=10per group.

[0028]FIG. 6. Role of peripheral sensory fibers inoleoylethanolamide-induced anorexia. Effects of vehicle (V),oleoylethanolamide (oleoylethanolamide/pranamide/OEA) (5 mg per kg,i.p.), CCK-8 (10 μg per kg) and CP-93129 (1 mg per kg), a centrallyactive 5-HT_(1B) receptor agonist, on food intake in a, control rats andc, capsaicin-treated rats. Water intake in (b) control rats and (d)capsaicin-treated rats. Asterisk, P<0.05; n=8-12 per group.

[0029]FIG. 7. Oleoylethanolamide/pranamide increases c-fos mRNAexpression in discrete brain regions associated with energy homeostasisand feeding behavior: (a) pseudocolor images of film autoradiographsshow that oleoylethanolamide (right section) elicits a striking andselective increase in c-fos mRNA labeling in the paraventricular (PVN)and supraoptic (SO) hypothalamic nuclei, as assessed by in situhybridization. A representative section from a vehicle-treated rat isshown at left. Labeling densities are indicated by color:blue<green<yellow<red. (b) quantification of c-fos cRNA labeling inforebrain regions [PVN, SO, arcuate (Arc), layer II piriform cortex(pir), ventrolateral thalamas (VI) and SI forelimb cortex (S 1FL)] ofrats treated with vehicle, oleoylethanolamide and oleic acid; (c) filmautoradiogram showing elevated ³⁵S c-fos mRNA expression in the nucleusof the solitary tract (NST) in an oleoylethanolamide-treated rat; Inset,c-fos cRNA labeling in the NST (shown in red) was identified by itslocalization relative to adjacent efferent nuclei (hypoglossal anddorsal motor nucleus of the vagus), which express choline acetyltransferase (ChAT) mRNA (shown in purple); (d) oleoylethanolamideincreases c-fos mRNA expression in NST but not in the hypoglossalnucleus (HgN). Two asterisks, P<0.0001, n=5 per group.

[0030]FIG. 8. The effects of OEA, Oleic acid (OA), AEA, PEA, andmethyl-OEA on fatty acid oxidation in soleus muscle.

DETAILED DESCRIPTION OF THE INVENTION

[0031] This invention relates to the surprising discovery that OEA andother fatty acid alkanolamide compounds act to reduce food intake, bodyweight, and body fat and to modulate fatty acid oxidation. It has beensurprisingly discovered that oleoylethanolamide (OEA), a natural lipidof heretofore unknown biological function in mammals, is a potent bodyfat reducing and weight control compound when administered to testanimals. U.S. Patent Application No. 60/279,542, filed Mar. 27, 2001,and assigned to the same assignee and herein incorporated by referencein its entirety discloses OEA and OEA-like compounds as agents which canreduce body fat and appetite in mammals.

[0032] Upon the discovery of the prototype OEA, other fatty acidalkanolamide compounds and homologs were also found to be active.

[0033] OEA can serve as a model in the development of other fatty acidalkanolamide-like fat reducing compounds for treating obesity, inducingweight loss, reducing appetite, or food intake. This invention providessuch other compounds as disclosed below.

[0034] The discovery that OEA adminstration acts to reduce appetite,food intake, and body weight can be used to identify other fatty acidethanolamides, homologues, and analogs as weight and appetite controlagents. This invention provides such agents.

Definitions

[0035] The abbreviations used herein have their conventional meaningwithin the chemical and biological arts.

[0036] Where substituent groups are specified by their conventionalchemical formulae, written from left to right, they equally encompassthe chemically identical substituents which would result from writingthe structure from right to left, e.g., —CH₂O— is intended to alsorecite —OCH₂—.

[0037] The term “composition”, as in pharmaceutical composition, isintended to encompass a product comprising the active ingredient(s), andthe inert ingredient(s) that make up the carrier, as well as any productwhich results, directly or indirectly, from combination, complexation oraggregation of any two or more of the ingredients, or from dissociationof one or more of the ingredients, or from other types of reactions orinteractions of one or more of the ingredients. Accordingly, thepharmaceutical compositions of the present invention encompass anycomposition made by admixing a compound of the present invention and apharmaceutically acceptable carrier. The term “pharmaceuticalcomposition” indicates a composition suitable for pharmaceutical use ina subject, including an animal or human. A pharmaceutical compositiongenerally comprises an effective amount of an active agent and apharmaceutically acceptable carrier.

[0038] Compounds of the invention may contain one or more asymmetriccenters and can thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Thepresent invention is meant to comprehend all such isomeric forms of theinventive compounds.

[0039] Some of the compounds described herein contain olefinic doublebonds, and unless specified otherwise, are meant to include both E and Zgeometric isomers.

[0040] Some of the compounds described herein may exist with differentpoints of attachment of hydrogen, referred to as tautomers. Such anexample may be a ketone and its enol form known as keto-enol tautomers.The individual tautomers as well as mixture thereof are encompassed bythe inventive formulas.

[0041] Compounds of the invention include the diastereoisomers of pairsof enantiomers. Diastereomers for example, can be obtained by fractionalcrystallization from a suitable solvent, for example methanol or ethylacetate or a mixture thereof. The pair of enantiomers thus obtained maybe separated into individual stereoisomers by conventional means, forexample by the use of an optically active acid as a resolving agent.

[0042] Alternatively, any enantiomer of an inventive compound may beobtained by stereospecific synthesis using optically pure startingmaterials or reagents of known configuration

[0043] As used herein, the term “heteroatom” is meant to include oxygen(O), nitrogen (N), sulfur (S) and silicon (Si).

[0044] “Alkanol,” as used herein refers to a saturated or unsaturated,substituted or unsubstituted, branched or unbranched alkyl group havinga hydroxyl substituent, or a substituent derivable from a hydroxylmoiety, e.g, ether, ester. The alkanol is preferably also substitutedwith a nitrogen-, sulfur-, or oxygen-bearing substituent that isincluded in bond Z (Formula I), between the “fatty acid” and thealkanol.

[0045] “Fatty acid,” as used herein, refers to a saturated orunsaturated substituted or unsubstituted, branched or unbranched alkylgroup having a carboxyl substituent. Preferred fatty acids are C₄-C₂₂acids. Fatty acid also encompasses species in which the carboxylsubstituent is replaced with a —CH₂— moiety.

[0046] The term “alkyl,” by itself or as part of another substituent,means, unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e. C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude, but are not limited to, groups such as methyl, ethyl, n-propyl,isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, forexample, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Anunsaturated alkyl group is one having one or more double bonds or triplebonds. Examples of unsaturated alkyl groups include, but are not limitedto, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “alkyl,” unlessotherwise noted, is also meant to include those derivatives of alkyldefined in more detail below, such as “heteroalkyl.” Alkyl groups whichare limited to hydrocarbon groups are termed “homoalkyl”.

[0047] The term “alkylene” by itself or as part of another substituentmeans a divalent radical derived from an alkane, as exemplified, but notlimited, by —CH₂CH₂CH₂CH₂—, and further includes those groups describedbelow as “heteroalkylene.” Typically, an alkyl (or alkylene) group willhave from 1 to 24 carbon atoms, with those groups having 10 or fewercarbon atoms being preferred in the present invention. A “lower alkyl”or “lower alkylene” is a shorter chain alkyl or alkylene group,generally having eight or fewer carbon atoms.

[0048] The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy)are used in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

[0049] The term “heteroalkyl,” by itself or in combination with anotherterm, means, unless otherwise stated, a stable straight or branchedchain, or cyclic hydrocarbon radical, or combinations thereof,consisting of the stated number of carbon atoms and at least oneheteroatom selected from the group consisting of O, N, Si and S, andwherein the nitrogen and sulfur atoms may optionally be oxidized and thenitrogen heteroatom may optionally be quaternized. The heteroatom(s) O,N and S and Si may be placed at any interior position of the heteroalkylgroup or at the position at which the alkyl group is attached to theremainder of the molecule. Examples include, but are not limited to,—CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃,—CH₂—CH₂, —S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃,—CH₂—CH═N—OCH₃, and —CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may beconsecutive, such as, for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.Similarly, the term “heteroalkylene” by itself or as part of anothersubstituent means a divalent radical derived from heteroalkyl, asexemplified, but not limited by, —CH₂—CH₂—S—CH₂—CH₂— and—CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylene groups, heteroatoms can alsooccupy either or both of the chain termini (e.g., alkyleneoxy,alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Stillfurther, for alkylene and heteroalkylene linking groups, no orientationof the linking group is implied by the direction in which the formula ofthe linking group is written. For example, the formula —C(O)₂R′—represents both —C(O)₂R′— and —R′C(O)₂—.

[0050] The terms “cycloalkyl” and “heterocycloalkyl”, by themselves orin combination with other terms, represent, unless otherwise stated,cyclic versions of “alkyl” and “heteroalkyl”, respectively.Additionally, for heterocycloalkyl, a heteroatom can occupy the positionat which the heterocycle is attached to the remainder of the molecule.Examples of cycloalkyl include, but are not limited to, cyclopentyl,cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like.Examples of heterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

[0051] The terms “halo” or “halogen,” by themselves or as part ofanother substituent, mean, unless otherwise stated, a fluorine,chlorine, bromine, or iodine atom. Additionally, terms such as“haloalkyl,” are meant to include monohaloalkyl and polyhaloalkyl. Forexample, the term “halo(C₁-C₄)alkyl” is mean to include, but not belimited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

[0052] The term “aryl” means, unless otherwise stated, apolyunsaturated, aromatic, hydrocarbon substituent which can be a singlering or multiple rings (preferably from 1 to 3 rings) which are fusedtogether or linked covalently. The term “heteroaryl” refers to arylgroups (or rings) that contain from one to four heteroatoms selectedfrom N, O, and S, wherein the nitrogen and sulfur atoms are optionallyoxidized, and the nitrogen atom(s) are optionally quaternized. Aheteroaryl group can be attached to the remainder of the moleculethrough a heteroatom. Non-limiting examples of aryl and heteroarylgroups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl,2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituentsfor each of the above noted aryl and heteroaryl ring systems areselected from the group of acceptable substituents described below.

[0053] For brevity, the term “aryl” includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like) including thosealkyl groups in which a carbon atom (e.g., a methylene group) has beenreplaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like).

[0054] Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl” and“heteroaryl”) are meant to include both substituted and unsubstitutedforms of the indicated radical. Preferred substituents for each type ofradical are provided below.

[0055] Substituents for the alkyl and heteroalkyl radicals (includingthose groups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′,-halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″,—NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and—NO₂ in a number ranging from zero to (2m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″, R′″ and R″″ eachpreferably independently refer to hydrogen, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted aryl, e.g., aryl substitutedwith 1-3 halogens, substituted or unsubstituted alkyl, alkoxy orthioalkoxy groups, or arylalkyl groups. When a compound of the inventionincludes more than one R group, for example, each of the R groups isindependently selected as are each R′, R″, R′″ and R″″ groups when morethan one of these groups is present. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant to include,but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the abovediscussion of substituents, one of skill in the art will understand thatthe term “alkyl” is meant to include groups including carbon atoms boundto groups other than hydrogen groups, such as haloalkyl (e.g., —CF₃ and—CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and thelike).

[0056] Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are varied and areselected from, for example: halogen, —OR′, ═O, ═NR′, ═N—OR′, —NR′R″,—SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —CN and —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy, andfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total number ofopen valences on the aromatic ring system; and where R′, R″, R′″ and R″″are preferably independently selected from hydrogen, (C₁-C₈)alkyl andheteroalkyl, unsubstituted aryl and heteroaryl, (unsubstitutedaryl)-(C₁-C₄)alkyl, and (unsubstituted aryl)oxy-(C₁-C₄)alkyl. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″and R″″ groups when more than one of these groups is present.

[0057] The term “body fat reduction” means loss of a portion of bodyfat.

[0058] The formula for Body Mass Index (BMI) is [Weight in pounds÷Heightin inches÷Height in inches]×703. BMI cutpoints for human adults are onefixed number, regardless of age or sex, using the following guidelines:Overweight human adults individuals have a BMI of 25.0 to 29.9. Obesehuman adults have a BMI of 30.0 or more. Underweight adults have a BMIless of than 18.5. A nomal body weight range for an adult is defined asa BMI between 18.5 and 25. BMI cutpoints for children under 16 aredefined according to percentiles: Overweight is defined as a BMI for agegreater than ≧85th percentile and obesity is defined as a BMI-for-age≧95th percentile. Underweight is a BMI-for-age <5th percentile. A normalbody weight range for a child is defined as a BMI above the 5thpercentile and below the 85 percentile.

[0059] The term “fatty acid oxidation” relates to the conversion offatty acids (e.g., oleate) into ketone bodies.

[0060] The term “hepatocytes” refers to cells originally derived fromliver tissue. Hepatocytes may be freshly isolated from liver tissue orestablished cell lines.

[0061] The term “modulate” means to induce any change includingincreasing or decreasing. (e.g., a modulator of fatty acid oxidationincreases or decreases the rate of fatty oxidation.

[0062] The term “muscle cells” refers to cells derived from thepredominant cells of muscle tissue. Muscle cells may be freshly isolatedfrom muscle tissue or established cell lines.

[0063] The term “obese” indicates a body weight 20% over ideal bodyweight as measured by body mass index

[0064] Oleoylethanolamide (OEA) refers to a natural lipid of thefollowing structure:

[0065] In the formulas herein, “Me” represents the methyl group.

[0066] The term “weight loss” refers to loss of a portion of total bodyweight.

[0067] The term “pharmaceutically acceptable carrier” encompasses any ofthe standard pharmaceutical carriers, buffers and excipients, includingphosphate-buffered saline solution, water, and emulsions (such as anoil/water or water/oil emulsion), and various types of wetting agentsand/or adjuvants. Suitable pharmaceutical carriers and theirformulations are described in REMINGTON'S PHARMACEUTICAL SCIENCES (MackPublishing Co., Easton, 19th ed. 1995). Preferred pharmaceuticalcarriers depend upon the intended mode of administration of the activeagent. Typical modes of administration are described below.

[0068] The term “effective amount” means a dosage sufficient to producea desired result. The desired result may comprise a subjective orobjective improvement in the recipient of the dosage. A subjectiveimprovement may be decreased appetite or craving for food. An objectiveimprovement may be decreased body weight, body fat, or food, decreasedfood consumption, or decreased food seeking behavior.

[0069] A “prophylactic treatment” is a treatment administered to asubject who does not exhibit signs of a disease or exhibits only earlysigns of a disease, wherein treatment is administered for the purpose ofdecreasing the risk of developing a pathology associated with increasedbody weight or body fat. The compounds of the invention may be given asa prophylactic treatment to prevent undesirable or unwanted weight gain.

[0070] A “therapeutic treatment” is a treatment administered to asubject who exhibits signs of pathology, wherein treatment isadministered for the purpose of diminishing or eliminating thosepathological signs.

[0071] The term “to control weight” encompasses the loss of body mass orthe reduction of weight gain over time.

[0072] The methods, compounds and compositions of the present inventionare generally useful for reducing or controlling body fat and bodyweight in mammals. For instance, the methods, compositions, andcompounds of the present invention are helpful in reducing appetite orinducing hypophagia in mammals. The methods, compounds, and compositionsare also useful in preventing or mitigating the diseases associated withoverweight or obesity by promoting the loss of body fat and body weight.

[0073] The methods, compositions, and compounds of the present inventioninclude modulators of lipid metabolism, and particularly, fat and fattyacid catabolism.

[0074] Compounds of the Invention

[0075] Certain compounds of the present invention may possess asymmetriccarbon atoms (optical centers) or double bonds; the racemates,diastereomers, geometric isomers and individual isomers are all intendedto be encompassed within the scope of the present invention.

[0076] Such compounds of the invention may be separated intodiastereoisomeric pairs of enantiomers by, for example, fractionalcrystallization from a suitable solvent, for example methanol or ethylacetate or a mixture thereof. The pair of enantiomers thus obtained maybe separated into individual stereoisomers by conventional means, forexample by the use of an optically active acid as a resolving agent.

[0077] Alternatively, any enantiomer of such a compound of the inventionmay be obtained by stereospecific synthesis using optically purestarting materials of known configuration.

[0078] The compounds of the present invention may have unnatural ratiosof atomic isotopes at one or more of their atoms. For example, thecompounds may be radiolabeled with isotopes, such as tritium orcarbon-14. All isotopic variations of the compounds of the presentinvention, whether radioactive or not, are within the scope of thepresent invention.

[0079] The instant compounds may be isolated in the form of theirpharmaceutically acceptable acid addition salts, such as the saltsderived from using inorganic and organic acids. Such acids may includehydrochloric, nitric, sulfuric, phosphoric, formic, acetic,trifluoroacetic, propionic, maleic, succinic, malonic and the like. Inaddition, certain compounds containing an acidic function can be in theform of their inorganic salt in which the counterion can be selectedfrom sodium, potassium, lithium, calcium, magnesium and the like, aswell as from organic bases. The term “pharmaceutically acceptable salts”refers to salts prepared from pharmaceutically acceptable non-toxicbases or acids including inorganic bases or acids and organic bases oracids.

[0080] The invention also encompasses prodrugs of the present compounds,which on administration undergo chemical conversion by metabolicprocesses before becoming active pharmacological substances. In general,such prodrugs will be derivatives of the present compounds that arereadily convertible in vivo into a functional compound of the invention.Conventional procedures for the selection and preparation of suitableprodrug derivatives are described, for example, in “Design of Prodrugs”,ed. H. Bundgaard, Elsevier, 1985. The invention also encompasses activemetabolites of the present compounds.

[0081] A. Fatty Acid Alkanolamide Compounds, Homologs, and Analogs.

[0082] Compounds of the invention include body fat reducing fatty acidalkanolamide compounds, including the fatty acid ethanolamide compounds,and their homologues and certain analogs of the fatty acidalkanolamides. Such compounds may be identified and defined in terms ofeither an ability to cause reduced appetite, food intake, and/or bodyweight or body fat upon administration to test animals in vivo.

[0083] A variety of such fatty acid alkanolamides, homologs and analogsare therefore contemplated. Compounds of the invention include compoundsof the following general formula:

[0084] In this formula, n is from 0 to 5 and the sum of a and b can befrom 0 to 4. Z is a member selected from —C(O)N(R^(o))—; —(R^(o))NC(O)—;—OC(O)—; —(O)CO—; O; NR^(o); and S, in which R^(o) and R² areindependently selected from the group consisting of unsubstituted orunsubstituted alkyl, hydrogen, substituted or unsubstituted C₁-C₆ alkyl,substituted or unsubstituted lower (C₁-C₆) acyl, homoalkyl, and aryl. Upto four hydrogen atoms of either or both the fatty acid portion andalkanolamine (e.g. ethanolamine) portion of the compound may also besubstituted by methyl or a double bond. In addition, the molecular bondbetween carbons c and d may be unsaturated or saturated. In someembodiments, the fatty acid ethanolamide of the above formula is anaturally occurring compound.

[0085] Compounds of the invention also include compounds of thefollowing formula:

[0086] In one embodiment, the compounds of Formula Ia have n from 0 to5; and a sum of a and b that is from 0 to 4; and members R¹ and R²independently selected from the group consisting of hydrogen,substituted or unsubstituted C₁-C₆ alkyl, lower substituted orunsubstituted (C₁-C₆) acyl, homoalkyl, and substituted or unsubstitutedaryl. In this embodiment, up to four hydrogen atoms of the fatty acidportion and alkanolamine (e.g., ethanolamine) portion of compounds ofthe above formula may also be substituted by methyl or a double bond. Inaddition, the molecular bond between carbons c and d may be unsaturatedor saturated. In some embodiments with acyl groups, the acyl groups maybe the propionic, acetic, or butyric acids and attached via an esterlinkage as R² or an amide linkage as R¹.

[0087] In another embodiment, the above compounds particularly includethose in which the fatty acid moiety comprises oleic acid, elaidic acid,or palmitic acid. Such compounds include oleoylethanolamide,elaidylethanolamide and palmitylethanolamide.

[0088] In another embodiment, the compounds of Formula Ia have n from 1to 3; and a sum of a and b that is from 1 to 3; and members R¹ and R²independently selected from the group consisting of hydrogen,substituted or unsubstituted C₁-C₆ alkyl, and lower substituted orunsubstituted (C₁-C₆) acyl. In this embodiment, up to four hydrogenatoms of the fatty acid portion and alkanolamine (e.g., ethanolamine)portion of compounds of the above formula may also be substituted bymethyl or a double bond. In addition, the molecular bond between carbonsc and d may be unsaturated or saturated. In a further embodiment, themolecular bond between carbons c and d is unsaturated and no otherhydrogen atoms are substituted. In a still further embodiment thereof,the members R¹ and R² are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₃ alkyl, andsubstituted or unsubstituted lower (C₁-C₃) acyl.

[0089] Exemplary compounds provide mono-methyl substituted compounds,including ethanolamides, of Formula Ia. Such compounds include:

[0090] The methyl substituted compounds of the above formula includeparticularly those compounds where R¹ and R² are both H:(R)1′-methyloleoylethanolamide, S(1′)-methyloleoylethanolamide,(R)2′-methyloleoylethanolamide, (S)2′-methyloleoylethanolamide,(R)1-methyloleoylethanolamide, and (S)1-methyloleoylethanolamide.

[0091] Reverse OEA-Like Compounds.

[0092] Compounds of the invention also include a variety of analogs ofOEA. These compounds include reverse OEA compounds of the generalformula:

[0093] In some embodiments, the invention provides compounds of FormulaII. Exemplary the compounds of Formula II have n from 1 to 5, and a sumof a and b from 0 to 4. In this embodiment, the member R² is selectedfrom the group consisting of hydrogen, substituted or unsubstitutedC₁-C₆ alkyl, substituted or unsubstituted lower (C₁-C₆) acyl, homoalkyl,and aryl. In addition, up to four hydrogen atoms of either or both thefatty acid portion and alkanolamine (e.g., ethanolamine) portion ofcompounds of the above formula may also be substituted by methyl or adouble bond.

[0094] Exemplary compounds of formula II include those compounds wherethe alkanolamine portion is ethanolamine, compounds where R² is H, andcompounds where a and b are each 1, and compounds where n is 1.

[0095] One embodiment of a compound according to Formula II is

[0096] In another embodiment, the compounds of Formula II have n from 1to 5 and a sum of a and b from 1 to 3. In this embodiment, the member R²is selected from the group consisting of hydrogen, substituted orunsubstituted C₁-C₆ alkyl, and substituted or unsubstituted lower(C₁-C₆) acyl. In addition, up to four hydrogen atoms of either or boththe fatty acid portion and alkanolamine (e.g., ethanolamine) portion ofcompounds of the above formula may also be substituted by methyl or adouble bond.

[0097] Oleoylalkanol Ester Compounds.

[0098] Compounds of the invention also include oleoylalkanol esters ofthe general formula:

[0099] In some embodiments, the compounds of Formula III, have n from 1to 5; and the sum of a and b from 0 to 4. The member R² is selected fromthe group consisting of hydrogen, substituted or unsubstituted C₁-C₆alkyl, lower (C₁-C₆) acyl, homoalkyl, and aryl. Up to four hydrogenatoms of either or both the fatty acid portion and alkanol (e.g.,ethanol) portion of compounds of the above formula may also besubstituted by methyl or a double bond.

[0100] In some embodiments, the compounds of Formula III, have n from 1to 3; and the sum of a and b from 1 to 3. The member R² is selected fromthe group consisting of hydrogen, substituted or unsubstituted C₁-C₆alkyl, and substituted or unsubstituted lower (C₁-C₆) acyl. Up to fourhydrogen atoms of the fatty acid portion and alkanol (e.g., ethanol)portion of compounds of the above formula may also be substituted bymethyl or a double bond.

[0101] Compounds of Formula III include those compounds where R² is H,compounds where a and b are each 1, and compounds where n is 1. Examplesof compounds according to Formula III include the oleoyldiethanol ester:

[0102] Compounds of Formula III also include mono-methyl substitutedoleoyl ethanol esters such as the (R or S)-2′-methyloleoylethanolesters;the (R or S)-1′-methyloleoylethanolesters; and the (R orS))-1′-methyloleoylethanolesters; respectively:

[0103] Oleoyl Alkanol Ethers

[0104] Compounds of the invention also include oleoylalkanol ethersaccording to the general formula:

[0105] In some embodiments, the compounds of Formula IV, have an n from1 to 5 and a sum of a and b that can be from 0 to 4. The member R² isselected from the group consisting of hydrogen, substituted orunsubstituted C₁-C₆ alkyl, substituted or unsubstituted lower (C₁-C₆)acyl, alkyl, and substituted and unsubstituted aryl. Up to four hydrogenatoms of either or both the fatty acid portion and alkanol (e.g.,ethanol) portion of compounds of the above formula may also besubstituted by methyl or a double bond.

[0106] In other embodiments, the compounds of Formula IV, have n from 1to 3; and the sum of a and b can be from 1 to 3. The member R² isselected from the group consisting of hydrogen, substituted orunsubstituted C₁-C₆ alkyl, and substituted or unsubstituted lower(C₁-C₆) acyl. Up to four hydrogen atoms of either or both the fatty acidportion and alkanol (e.g., ethanol) portion of compounds of the aboveformula may also be substituted by methyl or a double bond.

[0107] Compounds of Formula IV include those compounds where R² is H,compounds where a and b are each 1, and compounds where n is 1. Examplesof compounds according to Formula IV include the following (R or S)1′-oleoylethanol ethers and (R or S)-2′-oleoylethanol ethers:

[0108] Fatty Acid Alkanolamide Analogs Having Polar Head Variants.

[0109] Compounds of the invention also include a variety of polar headanalogs of OEA. These compounds include compounds having a fatty acidmoiety of the general formula:

[0110] In some embodiments, the compounds of Formula V have a sum of aand b that can be from 0 to 4. In other embodiments, the sum of a and bis from 1 to 3. In these embodiments, up to four hydrogen atoms of thecompounds of the above formula may also be substituted by methyl or adouble bond. In addition, the molecular bond between carbons c and d maybe unsaturated or saturated. A particularly preferred embodiment is thatof the oleic acid fatty acid moiety:

[0111] The R³ group of the above structures may be selected from any ofthe following:

[0112] HO-(CH₂)_(z)—NH— wherein z is from 1 to 5, and the alkyl portionthereof is an unbranched methylene chain. For example:

[0113] H₂N—(CH₂)_(z)—NH— wherein z is from 1 to 5, and the alkyl portionthereof is an unbranched methylene chain. For example:

[0114] HO—(CH₂)_(x)—NH— wherein x is from 1 to 8, and the alkyl portionthereof may be branched or cyclic. For example,

[0115] Additional polar head groups for R³ include, for instance,compounds having furan, dihydrofuran and tetrahydrofuran functionalgroups:

[0116] In the above structures, z can be from 1 to 5.

[0117] Compounds of the invention include, for instance, those having R³polar head groups based upon pyrole, pyrrolidine, and pyrroline rings:

[0118] In the compounds of the above structures, z can be from 1 to 5.

[0119] Other exemplary polar head groups includea variety of imidazoleand oxazoles, for example:

[0120] In the compounds of the above structures, z can be from 1 to 5.

[0121] Oxazolpyridine polar head groups are also exemplary:

[0122] Fatty Acid Alkanolamide Analogs Having Apolar Tail Variants.

[0123] Compounds of the invention include a variety of alkanolamide andethanolamide compounds having a variety of flexible apolar tails. Thesecompounds include compounds of the following formulas in which Rrepresents an ethanolamine moiety, an alkanolamine moiety, or a stableanalog thereof. In the case of ethanolamine, the ethanolamine moiety isattached preferably via the ethanolamine nitrogen rather than theethanolamine oxygen.

[0124] In the above structures, m is from 1 to 9 and p is independentlyfrom 1 to 5.

[0125] An exemplary compound is:

[0126] Another exemplary compound is an ethanolamine analog with anapolar tail of the following structural formula:

[0127] Exemplary compounds include analogs of fatty acid alkanolamides.Such analogs include those compounds taught in U.S. Pat. No. 6,200,998(hereby incorporated by reference). This reference teaches compounds ofthe general formula:

[0128] In the above formula, and as defined in U.S. Pat. No. 6,200,998,Ar¹ is (1) arylene or (2) heteroarylene, wherein arylene andheteroarylene are optionally substituted with from 1 to 4 groupsselected from R^(a); Ar² is (1) ortho-substituted aryl or (2)ortho-substituted heteroaryl, wherein said ortho substituent is selectedfrom R; and aryl and heteroaryl are optionally further substituted withfrom 1-4 groups independently selected from R^(a); X and Y areindependently O, S, N—R^(b), or CH₂; Z is O or S; n is 0 to 3; R is (1)C₃₋₁₀ alkyl optionally substituted with 1-4 groups selected from haloand C₃₋₆ cycloalkyl, (2) C₃₋₁₀ alkenyl, or (3) C₃₋₈ cycloalkyl; R^(a) is(1) C₁₋₁₅ alkanoyl, (2) C₁₋₁₅ alkyl, (3) C₂₋₁₅ alkenyl, (4) C₂₋₁₅alkynyl, (5) halo, (6) OR^(b), (7) aryl, or (8) heteroaryl, wherein saidalkyl, alkenyl, alkynyl, and alkanoyl are optionally substituted withfrom 1-5 groups selected from R^(c), and said aryl and heteroaryloptionally substituted with 1 to 5 groups selected from R^(d); R^(b) is(1) hydrogen, (2) C₁₋₁₀ alkyl, (3) C₂₋₁₀ alkenyl, (4) C₂₋₁₀ alkynyl, (5)aryl, (6) heteroaryl, (7) aryl C₁₋₁₅ alkyl, (8) heteroaryl C₁₋₁₅ alkyl,(9) C₁₋₁₅ alkanoyl, (10) C₃₋₈ cycloalkyl, wherein alkyl, alkenyl,alkynyl are optionally substituted with one to four substituentsindependently selected from R^(c), and cycloalkyl, aryl and heteroarylare optionally substituted with one to four substituents independentlyselected from R^(d); or R^(c) is (1) halo, (2) aryl, (3) heteroaryl, (4)CN, (5) NO₂, (6) OR^(f); (7) S(O)_(m)R^(f), m=0, 1 or 2, provided thatR^(f) is not H when m is 1 or 2; (8) NR^(f)R^(f), (9) NR^(f)COR^(f),(10) NR^(f)CO₂ R^(f), (11) NR^(f)CON(R^(f))₂, (12) NR^(f) SO₂ R^(f),provided that R^(f) is not H, (13) COR^(f), (14) CO₂R^(f), (15)CON(R^(f))₂, (16) SO₂ N(R^(f))₂, (17) OCON(R^(f))₂, or (18) C₃₋₈cycloalkyl, wherein said cycloalkyl, aryl and heteroaryl are optionallysubstituted with 1 to 3 groups of halo or C₁₋₆ alkyl; R^(d) is (1) agroup selected from R^(c), (2) C₁₋₁₀ alkyl, (3) C₂₋₁₀ alkenyl, (4) C₂₋₁₀alkynyl, (5) aryl C₁₋₁₀ alkyl, or (6) heteroaryl C₁₋₁₀ alkyl, whereinalkyl, alkenyl, alkynyl, aryl, heteroaryl are optionally substitutedwith a group independently selected from R^(e); R^(e) is (1) halogen,(2) amino, (3) carboxy, (4) C₁₋₄ alkyl, (5) C₁₋₄ alkoxy, (6) hydroxy,(7) aryl, (8) aryl C₁₋₄ alkyl, or (9) aryloxy; R^(f) is (1) hydrogen,(2) C₁₋₁₀ alkyl, (3) C₂₋₁₀ alkenyl, (4) C₂₋₁₀ alkynyl, (5) aryl, (6)heteroaryl, (7) aryl C₁₋₁₅ alkyl, (8) heteroaryl C₁₋₁₅ alkyl, (9) C₁₋₁₅alkanoyl, (10) C₃₋₈ cycloalkyl; wherein alkyl, alkenyl, alkynyl, aryl,heteroaryl, alkanoyl and cycloalkyl are optionally substituted with oneto four groups selected from R^(e).

[0129] Also preferred are the analogs taught in U.S. Pat. No. 5,859,051.These analogs have the following general formula:

[0130] In the embodiments according to Formula VII, as defined in U.S.Pat. No. 5,859,051, R¹ is selected from the group consisting of H, C₁₋₆alkyl, C₅₋₁₀ aryl, and C₅₋₁₀ heteroaryl, said alkyl, aryl and heteroaryloptionally substituted with 1 to 3 groups of R^(a); R¹ is selected froma group consisting of: H, C₁₋₁₅ alkyl, C₂₋₁₅ alkenyl, C₂₋₁₅ alkynyl andC₃₋₁₀ cycloalkyl, said alkyl, alkenyl, alkynyl, and cycloalkyloptionally substituted with 1 to 3 groups of R^(a); R³ is selected froma group consisting of: H, NHR¹, NHacyl, C₁₋₁₅ alkyl, C₃₋₁₀ cycloalkyl,C₂₋₁₅ alkenyl, C₁₋₁₅ alkoxy, CO₂ alkyl, OH, C₂₋₁₅ alkynyl, C₅₋₁₀ aryl,C₅₋₁₀ heteroaryl said alkyl, cycloalkyl, alkenyl, alkynyl, aryl andheteroaryl optionally substituted with 1 to 3 groups of R^(a); (Z—W—) isZ—CR⁶R⁷—, Z—CH.═CH—, or:

[0131] R⁸ is selected from the group consisting of CR⁶R⁷, O, NR.⁶, andS(O)_(P); R⁶ and R⁷ are independently selected from the group consistingof H, C₁₋₆ alkyl; B is selected from the group consisting of: 1) a 5 or6 membered heterocycle containing 0 to 2 double bonds, and 1 heteroatomselected from the group consisting of O, S and N, heteroatom beingsubstituted at any position on the five or six membered heterocycle, theheterocycle being optionally unsubstituted or substituted with 1 to 3groups of R^(a); 2) a 5 or 6 membered carbocycle containing 0 to 2double bonds, the carbocycle optionally unsubstituted or substitutedwith 1 to 3 groups of R^(a) at any position on the five or six memberedcarbocycle; and 3) a 5 or 6 membered heterocycle containing 0 to 2double bonds, and 3 heteroatoms selected from the group consisting of O,N, and S, which are substituted at any position on the five or sixmembered heterocycle, the heterocycle being optionally unsubstituted orsubstituted with 1 to 3 groups of R^(a); X¹ and X² are independentlyselected from a group consisting of: H, OH, C₁₋₁₅ alkyl, C₂₋₁₅ alkenyl,C₂₋₁₅ alkynyl, halo, OR³, ORCF₃, C₅₋₁₀ aryl, C₅₋₁₀ aralkyl, C₅₋₁₀heteroaryl and C₁₋₁₀ acyl, said alkyl, alkenyl, alkynyl, aryl andheteroaryl optionally substituted with 1 to 3 groups of R^(a); R^(a)represents a member selected from the group consisting of: halo, acyl,aryl, heteroaryl, CF₃, OCF₃, —O—, CN, NO₂, R³, OR³; SR³, ═N(OR), S(O)R³,SO₂R³, NR³R³, NR³ COR³, NR³ CO₂ R³, NR³CON(R³)₂, NR³ SO₂ R³, COR³,CO₂R³, CON(R³)₂, SO₂ N(R³)₂, OCON(R³)₂ said aryl and heteroaryloptionally substituted with 1 to 3 groups of halo or C₁₋₆ alkyl; Y isselected from the group consisting of: S(O)_(p), —CH₂—, —C(O)—,—C(O)NH—, —NR—, —O—, —SO₂NH—, —NHSO₂; Y¹ is selected from the groupconsisting of: O and C; Z is selected from the group consisting of:CO₂R³, R³CO₂R³, CONHSO₂Me, CONHSO₂, CONH₂ and 5-(1H-tetrazole); t and vare independently 0 or 1 such that t+v=1 Q is a saturated or unsaturatedstraight chain hydrocarbon containing 2-4 carbon atoms and p is 0-2 withthe proviso when Z is CO₂ R³ and B is a 5 membered heterocycleconsisting of 0, R³ does not represent methyl.

[0132] Additional analogs suitable for practicing the inventive methodsand compositions include compounds taught in U.S. Pat. Nos. 5,847,008,6,090,836 and 6,090,839, each of which is herein incorporated byreference in its entirety to the extent not inconsistent with thepresent disclosure.

[0133] Additionally a variety of suitable analogs are taught in U.S.Pat. No. 6,274,608. Aryl and heteroaryl acetic acid and oxyacetic acidanalogs are taught for instance in U.S. Pat. No. 6,160,000; substituted5-aryl-2,4-thiazolidinedione analogs are taught in U.S. Pat. No.6,200,998; other possible analogs such as polyunsaturated fatty acidsand eicosanoids are known (see for instance, Forman, B M, Chen, J, andEvans R M, PNAS 94:4312-4317. The compounds of these publications, whichare each herein incorporated by reference in their entirety to theextent not inconsistent with the present disclosure can be screened bythe methods provide below to provide compounds which are useful, forinstance, in reducing body fat. and body weight, modulating fatcatabolism, and reducing appetite according to the present disclosure.

[0134] Synthesis of Fatty Acid Alkanolamides

[0135] Compounds useful in practicing the present invention are readilysynthesized and purified using methods recognized in the art. In anexemplary synthetic scheme (Scheme 1), a carboxylic acid and anaminoalcohol (or an O-protected derivative thereof) are reacted in a thepresence of a dehydrating agent, e.g., dicyclohexylcarbodiimide, in anappropriate solvent. The fatty acid alkanol amide is isolated by methodssuch as extraction, crystallization, precipitation, chromatography andthe like. If the final product is the O-protected adduct, it isdeprotected, typically by an art-recognized method, to afford a fattyacid adduct having a free hydroxyl group.

[0136] Those of skill in the art will recognize that many variants onthe scheme set forth above are available. For example, an activatedderivative, e.g, acyl halide, active ester, of the acid can be used.Similarly, a glycol (preferably mono O-protected) can be substituted forthe amino alcohol, resulting in an ester linkage between the twoconstituents of the molecule.

[0137] Reverse esters and reverse amides are also readily synthesized byart-recognized methods. For example, a hydroxycarboxylic acid is reactedwith an amine or hydroxy derivative of a long chain alkyl (i.e., C₄-C₂₂)in the presence of a dehydrating agent. In certain reaction pathways, itis desirable to protect the hydroxyl moiety of the hydroxycarboxylicacid.

[0138] Ethers and mercaptans are prepared by methods well-known to thoseof skill in the art, e.g., Williamson synthesis. For example, a longchain alkyl alcohol or thiol is deprotonated by a base, e.g, NaH, and areactive alcohol derivative, e.g., a halo, tosyl, mesyl alcohol, or aprotected derivative thereof is reacted with the resulting anion to formthe ester or mercaptan.

[0139] The above-recited methods and variations thereof can be found in,for example, RECENT DEVELOPMENTS IN THE Synthesis OF FATTY ACIDDERIVATIVES, Knothe G, ed., Amer. Oil Chemists Society 1999;COMPREHENSIVE NATURAL PRODUCTS CHEMISTRY AND OTHER SECONDARY METABOLITESINCLUDING FATTY ACIDS AND THEIR DERIVATIVES, Nakanishi K, ed., PergamonPress, 1999; ORGANIC SYNTHESIS COLLECTED VOLUMES I-V, John Wiley andSons; COMPENDIUM OF ORGANIC SYNTHETIC METHODS, Volumes 1-6, WileyInterscience 1984; ORGANIC FUNCTIONAL GROUP PREPARATION, Volumes I-III,Academic Press Ltd. 1983; Greene T, PROTECTING GROUPS IN ORGANICSYNTHESIS, 2d ed., Wiley Interscience 1991.

[0140] Methods of use, Pharmaceutical Compositions, and theirAdministration

[0141] Methods of Use

[0142] The compounds, compositions and methods of the invention (e.g.,fatty acid alkanolamides, fatty acid ethanolamide compounds, analogs,and homologues) are used to reduce body fat and or body weight inmammals, including dogs, cats, and especially humans. The weight lossmay be for aesthetic or therapeutic purposes. The compounds may also beused to reduce appetite or induce hypophagia.

[0143] The compounds, compositions, and methods of the invention areused to prevent weight gain or body fat increases in individuals withina normal weight range. The compounds may be used in otherwise healthyindividuals who are not otherwise in need of any pharmaceuticalintervention for diseases related to diabetes or hyperlipidemia orcancer. In some embodiments, the individuals to be treated are free ofdiseases related to disturbances in sugar or lipid levels or metabolismor free of risk factors for cardiovascular and cerebrovascular disease.The individuals may be non-diabetic and have blood sugar levels in thenormal range. The individuals may also have blood lipids (e.g.,cholesterol) or triglyceride levels in the normal range. The individualsmay be free of atherosclerosis. The individuals may be free of otherconditions such as cancer or other tumors, disorders involving insulinresistance, Syndrome X, and pancreatitis.

[0144] In other embodiments, the subjects are overweight or obesepersons in need of body fat and/or body weight reduction. In theseembodiments, the methods, compounds, and compositions of the inventioncan be administered to promote weight loss and also to prevent weightgain once a body weight within the normal range for a person of that sexand age and height has been achieved. The compounds may be used inotherwise healthy individuals who are not in need of any pharmaceuticaltreatment of a disorder related to diabetes, hyperlipidemia, or cancer.The individuals may also otherwise free of risk factors forcardiovascular and cerebrovascular diseases. In some embodiments, theindividuals to be treated are free of diseases related to sugar (e.g.,glucose) or lipid metabolism. The individuals may be non-diabetic andhave blood sugar levels in the normal range. The individuals may alsohave blood lipids (e.g., cholesterol, HDL, LDL, total cholesterol) ortriglyceride levels in the normal range. The individuals may not need tobe in treatment for atherosclerosis.

[0145] The compounds methods, and compositions of the invention may alsobe administered to suppress appetite in mammals, including cats, dogs,and humans. In some embodiments, the compounds may be used in otherwisehealthy individuals who are not in need of pharmaceutical interventionsfor any disease. In some embodiments, the individuals do not needpreventive or ameliorative therapy for diseases, including cancer,diabetes, or hyperlipidemia. In some embodiments, the individuals to betreated are free of diseases related to abnormal sugar or lipid levels.In other embodiments the individuals may be free of risk factors forcardiovascular or cerebrovascular disease. The individuals may benon-diabetic and have blood sugar levels in the normal range. Theindividuals may also have blood lipids (e.g., cholesterol) ortriglyceride levels in the normal range. The individuals may be free ofatherosclerosis.

[0146] The compounds methods, and compositions of the invention may alsobe administered to modulate fat metabolism (e.g., increase fatcatabolism) in mammals, including cats, dogs, and humans. In someembodiments, the compounds may be used to reduce appetite in otherwisehealthy individuals. In some embodiments, the individuals to be treatedare free of diseases related to sugar or lipid metabolism (e.g.,diabetes, hypercholesterolemia, low HDL levels or high LDL levels). Theindividuals may be non-diabetic and have blood sugar levels in thenormal range. The individuals may also have blood lipids (e.g.,cholesterol) or triglyceride levels in the normal range. The individualsmay be free of atherosclerosis.

[0147] Treatment with the compounds and compositions of the inventionmay be for a period predetermined by the degree or amount of weight losshas been accomplished or when the individual achieves a BMI within thenormal range. Treatment with the compounds and compositions of theinvention may be reduced once a predetermined degree or amount of weightloss has been accomplished or when the individual achieves a BMI withinthe normal range

[0148] The compounds and compositions of the invention may beadministered solely for the purposes of reducing body fat or reducingappetite.

[0149] Pharmaceutical Compositions.

[0150] Another aspect of the present invention provides pharmaceuticalcompositions which comprise compounds of the invention and apharmaceutically acceptable carrier.

[0151] The pharmaceutical compositions of the present invention comprisea compound of the instant invention as an active ingredient or apharmaceutically acceptable salt thereof, and may also contain apharmaceutically acceptable carrier and optionally other therapeuticingredients.

[0152] The compositions include compositions suitable for oral, rectal,topical, parenteral (including subcutaneous, intramuscular, andintravenous), ocular (ophthalmic), pulmonary (nasal or buccalinhalation), or nasal administration, although the most suitable routein any given case will depend in part on the nature and severity of theconditions being treated and on the nature of the active ingredient. Anexemplary route of administration is the oral route. The compositionsmay be conveniently presented in unit dosage form and prepared by any ofthe methods well-known in the art of pharmacy.

[0153] In practical use, the compounds of the invention can be combinedas the active ingredient in intimate admixture with a pharmaceuticalcarrier according to conventional pharmaceutical compounding techniques.The carrier may take a wide variety of forms depending on the form ofpreparation desired for administration, e.g., oral or parenteral(including intravenous). In preparing the compositions for oral dosageform, any of the usual pharmaceutical media may be employed, such as,for example, water, glycols, oils, alcohols, flavoring agents,preservatives, coloring agents and the like in the case of oral liquidpreparations, such as, for example, suspensions, elixirs and solutions;or carriers such as starches, sugars, microcrystalline cellulose,diluents, granulating agents, lubricants, binders, disintegrating agentsand the like in the case of oral solid preparations such as, forexample, powders, hard and soft capsules and tablets, with the solidoral preparations being preferred over the liquid preparations.

[0154] Because of their ease of administration, tablets and capsulesrepresent the most advantageous oral dosage unit form in which casesolid pharmaceutical carriers are obviously employed. If desired,tablets may be coated by standard aqueous or nonaqueous techniques. Suchcompositions and preparations can contain at least 0.1 percent of activecompound. The percentage of active compound in these compositions may,of course, be varied and may conveniently be between about 2 percent toabout 60 percent of the weight of the unit. The amount of activecompound in such therapeutically useful compositions is such that atherapeutically effective dosage will be obtained. The active compoundscan also be administered intranasally as, for example, liquid drops orspray.

[0155] The tablets, pills, capsules, and the like may also contain abinder such as gum tragacanth, acacia, corn starch or gelatin;excipients such as dicalcium phosphate; a disintegrating agent such ascorn starch, potato starch, alginic acid; a lubricant such as magnesiumstearate; and a sweetening agent such as sucrose, lactose or saccharin.When a dosage unit form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier such as a fatty oil.

[0156] Various other materials may be present as coatings or to modifythe physical form of the dosage unit. For instance, tablets may becoated with shellac, sugar or both. A syrup or elixir may contain, inaddition to the active ingredient, sucrose as a sweetening agent, methyland propylparabens as preservatives, a dye and a flavoring such ascherry or orange flavor. To prevent breakdown during transit through theupper portion of the GI tract, the composition may be an enteric coatedformulation.

[0157] Administration

[0158] The compounds of the invention may also be administeredparenterally. Solutions or suspensions of these active compounds can beprepared in water suitably mixed with a surfactant such ashydroxypropylcellulose. Dispersions can also be prepared in glycerol,liquid polyethylene glycols and mixtures thereof in oils. Under ordinaryconditions of storage and use, these preparations contain a preservativeto prevent the growth of microorganisms.

[0159] The pharmaceutical forms suitable for injectable use includesterile aqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g. glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0160] The compounds of the invention can be effective over a widedosage range. For example, in the treatment of adult humans, dosagesfrom about 10 to about 1000 mg, about 100 to about 500 mg or about 1 toabout 100 mg may be needed. Doses of the 0.05 to about 100 mg, and morepreferably from about 0.1 to about 100 mg, per day may be used. A mostpreferable dosage is about 0.1 mg to about 70 mg per day. In choosing aregimen for patients, it may frequently be necessary to begin with adosage of from about 2 to about 70 mg per day and when the condition isunder control to reduce the dosage as low as from about 0.1 to about 10mg per day. For example, in the treatment of adult humans, dosages fromabout 0.05 to about 100 mg, preferably from about 0.1 to about 100 mg,per day may be used. The exact dosage will depend upon the mode ofadministration, on the therapy desired, form in which administered, thesubject to be treated and the body weight of the subject to be treated,and the preference and experience of the physician or veterinarian incharge.

[0161] Generally, the compounds of the present invention can bedispensed in unit dosage form comprising preferably from about 0.1 toabout 100 mg of active ingredient together with a pharmaceuticallyacceptable carrier per unit dosage. Usually, dosage forms suitable fororal, nasal, pulmonary or transdermal administration comprise from about0.001 mg to about 100 mg, preferably from about 0.01 mg to about 50 mgof the compounds admixed with a pharmaceutically acceptable carrier ordiluent. For storage and use, these preparations preferably contain apreservative to prevent the growth of microorganisms.

[0162] Administration of an appropriate amount the candidate compoundmay be by any means known in the art such as, for example, oral orrectal, parenteral, intraperitoneal, intravenous, subcutaneous,subdermal, intranasal, or intramuscular. In some embodiments,administration is transdermal. An appropriate amount or dose of thecandidate compound may be determined empirically as is known in the art.An appropriate or therapeutic amount is an amount sufficient to effect aloss of body fat or a loss in body weight in the animal over time. Thecandidate compound can be administered as often as required to effect aloss of body fat or loss in body weight, for example, hourly, every six,eight, twelve, or eighteen hours, daily, or weekly

[0163] Formulations suitable for oral administration can consist of (a)liquid solutions, such as an effective amount of the packaged nucleicacid suspended in diluents, such as water, saline or PEG 400; (b)capsules, sachets or tablets, each containing a predetermined amount ofthe active ingredient, as liquids, solids, granules or gelatin; (c)suspensions in an appropriate liquid; and (d) suitable emulsions. Tabletforms can include one or more of lactose, sucrose, mannitol, sorbitol,calcium phosphates, corn starch, potato starch, microcrystallinecellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate,stearic acid, and other excipients, colorants, fillers, binders,diluents, buffering agents, moistening agents, preservatives, flavoringagents, dyes, disintegrating agents, and pharmaceutically compatiblecarriers. Lozenge forms can comprise the active ingredient in a flavor,e.g., sucrose, as well as pastlles comprising the active ingredient inan inert base, such as gelatin and glycerin or sucrose and acaciaemulsions, gels, and the like containing, in addition to the activeingredient, carriers known in the art.

[0164] Injection solutions and suspensions can be prepared from sterilepowders, granules, and tablets of the kind previously described.Formulations suitable for parenteral administration, such as, forexample, by intraarticular (in the joints), intravenous, intramuscular,intradermal, intraperitoneal, and subcutaneous routes, include aqueousand non-aqueous, isotonic sterile injection solutions, which can containantioxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.

[0165] With respect to transdermal routes of administration, methods fortransdermal administration of drugs are disclosed in Remington'sPharmaceutical Sciences, 17th Edition, (Gennaro et al. Eds., MackPublishing Co., 1985). Dermal or skin patches are a preferred means fortransdermal delivery of the compounds of the invention. Patchespreferably provide an absorption enhancer such as DMSO to increase theabsorption of the compounds. Other methods for transdermal drug deliveryare disclosed in U.S. Pat. No. 5,962,012, 6,261,595, and 6,261,595. Eachof which is incorporated by reference in its entirety.

[0166] Preferred patches include those that control the rate of drugdelivery to the skin. Patches may provide a variety of dosing systemsincluding a reservoir system or a monolithic system, respectively. Thereservoir design may, for example, have four layers: the adhesive layerthat directly contacts the skin, the control membrane, which controlsthe diffusion of drug molecules, the reservoir of drug molecules, and awater-resistant backing. Such a design delivers uniform amounts of thedrug over a specified time period, the rate of delivery has to be lessthan the saturation limit of different types of skin.

[0167] The monolithic design, for example, typically has only threelayers: the adhesive layer, a polymer matrix containing the compound,and a water-proof backing. This design brings a saturating amount ofdrug to the skin. Thereby, delivery is controlled by the skin. As thedrug amount decreases in the patch to below the saturating level, thedelivery rate falls.

[0168] Compounds of the invention may be used in combination with othercompounds of the invention or with other drugs that may also be usefulin dieting or the treatment, prevention, suppression or amelioration ofbody fat. Such other drugs may be administered, by a route and in anamount commonly used therefor, contemporaneously or sequentially with acompound of the invention. When a compound of the invention is usedcontemporaneously with one or more other drugs, a pharmaceuticalcomposition in unit dosage form containing such other drugs and thecompound is preferred. When used in combination with one or more otheractive ingredients, the compound of the present invention and the otheractive ingredients may be used in lower doses than when each is usedsingly. Accordingly, the pharmaceutical compositions of the presentinvention include those that contain one or more other activeingredients, in addition to the compounds disclosed above.

[0169] Identification of Compounds of the Invention

[0170] Candidate compounds, such as disclosed above, can be screened bya variety of means known in the art. Body fat reducing compounds, forinstance, can be identified in vivo using animal bioassay techniquesknown to those of ordinary skill in the art. Test compounds andappropriate vehicle or caloric controls can be administered by any of anumber of routes (e.g., the oral route, a parenteral route) toexperimental subjects and the weight of the subjects can be monitoredover the course of therapy. The experimental subjects are humans or testanimals (e.g., rats, mice).

[0171] The effect of the compound on appetite or in inducing hypophagiaor reduced food intake can be assessed, for instance, by monitoring thefood consumption of the test subjects (e.g., measuring the amount eatenor not eaten by a subject in terms of food weight or caloric content).The effect of the compounds on appetite can also be assessed bysubjective means including questionnaires as to appetite or foodcravings levels by human subjects. The effect of the test compounds onlipid metabolism can be assessed by monitoring blood lipids and fattyacid oxidation. The techniques for these assessments are well known tothose of ordinary skill in the art. The studies may be acute, subacute,chronic, or subchronic with respect to the duration of administrationand or follow-up of the effects of the administration.

[0172] Body fat reduction can be determined, for instance, by directlymeasuring changes in body fat of the animal or by measuring changes inthe body weight of the animal. The animal may selected from the groupconsisting of a mouse, a rat, a guinea pig, or a rabbit. The animal mayalso be an ob/ob mouse, a db/db mouse, or a Zucker rat or other animalmodel for a weight-associated disease. Clinical studies in humans mayalso be conducted.

[0173] Combinatorial Chemical Libraries

[0174] Recently, attention has focused on the use of combinatorialchemical libraries to assist in the generation of new chemical compoundleads. A combinatorial chemical library is a collection of diversechemical compounds generated by either chemical synthesis or biologicalsynthesis by combining a number of chemical “building blocks” such asreagents. For example, a linear combinatorial chemical library such as apolypeptide library is formed by combining a set of chemical buildingblocks called amino acids in every possible way for a given compoundlength (i.e., the number of amino acids in a polypeptide compound).Millions of chemical compounds can be synthesized through suchcombinatorial mixing of chemical building blocks. For example, onecommentator has observed that the systematic, combinatorial mixing of100 interchangeable chemical building blocks results in the theoreticalsynthesis of 100 million tetrameric compounds or 10 billion pentamericcompounds (Gallop et al. J. Med. Chem. 37(9):1233(1994)).

[0175] Preparation and screening of combinatorial chemical libraries arewell known to those of skill in the art. Such combinatorial chemicallibraries include, but are not limited to, p benzodiazepines (U.S. Pat.No. 5,288,514), diversomers such as hydantoins, benzodiazepines anddipeptides (Hobbs et al. PNAS USA 90: 6909(1993)), analogous organicsyntheses of small compound libraries (Chen et al.) J. Amer. Chem. Soc.116: 2661(1994), oligocarbamates (Cho, et al., Science 261: 1303(1993)),and/or peptidyl phosphonates (Campbell et al., J. Org. Chem. 59:658(1994)), and small organic molecule libraries (see, e.g.,benzodiazepines (Baum C&EN, January 18, page 33(1993)), thiazolidinonesand metathiazanones (U.S. Pat. No. 5,549,974), pyrrolidines (U.S. Pat.Nos. 5,525,735 and 5,519,134), benzodiazepines (U.S. Pat. No.5,288,514), and the like.

[0176] Devices for the preparation of combinatorial libraries arecommercially available (see, e.g., 357 MPS, 390 MPS, Advanced Chem Tech,Louisville Ky., Symphony, Rainin, Woburn, Mass., 433A AppliedBiosystems, Foster City, Calif., 9050 Plus, Millipore, Bedford, Mass.).

[0177] A number of well known robotic systems have also been developedfor solution phase chemistries. These systems include automatedworkstations like the automated synthesis apparatus developed by TakedaChemical Industries, LTD. (Osaka, Japan) and many robotic systemsutilizing robotic arms (Zymate II, Zymark Corporation, Hopkinton, Mass.;Orca, HewlettPackard, Palo Alto, Calif.) which mimic the manualsynthetic operations performed by a chemist. Any of the above devicesare suitable for use with the present invention. The nature andimplementation of modifications to these devices so that they canoperate as discussed herein will be apparent to persons skilled in therelevant art. In addition, numerous combinatorial libraries arethemselves commercially available (see, e.g., ComGenex, Princeton, N.J.,Asinex, Moscow, Ru, Tripos, Inc., St. Louis, Mo., ChemStar, Ltd.,Moscow, RU, 3D Pharmaceuticals, Exton, Pa., Martek Biosciences,Columbia, Md., etc.).

[0178] High Throughput Assays of Chemical Libraries

[0179] The assays for compounds described herein are amenable to highthroughput screening. Preferred assays thus detect activation oftranscription (i.e., activation of mRNA production) by the testcompound(s), activation of protein expression by the test compound(s),or binding to the gene product (e.g., expressed protein) by the testcompound(s); or effects on fatty acid modulation as described below.

[0180] High throughput assays for the presence, absence, orquantification of particular protein products or binding assays are wellknown to those of skill in the art. Thus, for example, U.S. Pat. No.5,559,410 discloses high throughput screening methods for proteins, andU.S. Pat. Nos. 5,576,220 and 5,541,061 disclose high throughput methodsof screening for ligand/antibody binding.

[0181] In addition, high throughput screening systems are commerciallyavailable (see, e.g., Zymark Corp., Hopkinton, Mass.; Air TechnicalIndustries, Mentor, Ohio; Beckman Instruments, Inc. Fullerton, Calif.;Precision Systems, Inc., Natick, Mass., etc.). These systems typicallyautomate entire procedures including all sample and reagent pipetting,liquid dispensing, timed incubations, and final readings of themicroplate in detector(s) appropriate for the assay. These configurablesystems provide high throughput and rapid start up as well as a highdegree of flexibility and customization. The manufacturers of suchsystems provide detailed protocols the various high throughput. Thus,for example, Zymark Corp. provides technical bulletins describingscreening systems for detecting the modulation of gene transcription,ligand binding, and the like.

[0182] Determining Whether Compounds Affect Food Intake, Body Weight,Body Fat, Appetite, Food Seeking Behavior, or Modulate Fatty AcidOxidation

[0183] Compounds of the invention can be administered to an animal todetermine whether they affect food intake and body weight, body fat,appetite, food seeking behavior, or modulate modulator fatty acidoxidation.

[0184] Animals can be, for example, obese or normal guinea pigs, rats,mice, or rabbits. Suitable rats include, for example, Zucker rats.Suitable mice include, for example, normal mice, ALS/LtJ,C3.SW-H-^(2b)/SnJ, (NON/LtJ×NZO/HlJ)F1, NZO/HlJ, ALR/LtJ, NON/LtJ,KK.Cg-AALR/LtJ, NON/LtJ, KK.Cg-A^(y)/J, B6.HRS(BKS)-Cpe^(fat)/+,B6.129P2-Gck^(tm/Efr), B6.V-Lep^(ob), BKS.Cg-m+/+Lep^(rd)b, and C57BL/6Jwith Diet Induced Obesity.

[0185] Administration of an appropriate amount the candidate compoundmay be by any means known in the art such as, for example, oral orrectal, parenteral such as, for example, intraperitoneal, intravenous,subcutaneous, subdermal, intranasal, or intramuscular. Preferablyadministration may be intraperitoneal or oral. An appropriate effectiveamount of the candidate compound may be determined empirically as isknown in the art. An appropriate effective amount may be an amountsufficient to effect a loss of body fat or a loss in body weight orreduction in food consumption in the animal over time. The candidatecompound can be administered as often as required to effect a loss ofbody fat or loss in body weight, for example, hourly, every six, eight,twelve, or eighteen hours, daily, or weekly.

[0186] Formulations suitable for oral administration include (a) liquidsolutions, such as an effective amount of the candidate compoundsuspended in diluents, such as water, saline or PEG 400; (b) capsules,sachets or tablets, each containing a predetermined amount of the activeingredient, as liquids, solids, granules or gelatin; (c) suspensions inan appropriate liquid; and (d) suitable emulsions. Tablet forms includeone or more of lactose, sucrose, mannitol, sorbitol, calcium phosphates,corn starch, potato starch, microcrystalline cellulose, gelatin,colloidal silicon dioxide, talc, magnesium stearate, stearic acid, andother excipients, colorants, fillers, binders, diluents, bufferingagents, moistening agents, preservatives, flavoring agents, dyes,disintegrating agents, and pharmaceutically compatible carriers. Lozengeforms can comprise the active ingredient in a flavor, e.g., sucrose, aswell as pastilles comprising the active ingredient in an inert base,such as gelatin and glycerin or sucrose and acacia emulsions, gels, andthe like containing, in addition to the active ingredient, carriersknown in the art.

[0187] Injection solutions and suspensions can be prepared from sterilepowders, granules, and tablets of the kind previously described.Formulations suitable for parenteral administration, include, forexample, aqueous and non-aqueous, isotonic sterile injection solutions,which can contain antioxidants, buffers, bacteriostats, and solutes thatrender the formulation isotonic with the blood of the intendedrecipient, and aqueous and non-aqueous sterile suspensions that caninclude suspending agents, solubilizers, thickening agents, stabilizers,and preservatives.

[0188] The dose administered to the animal is sufficient to effect achange in body weight, body fat, and/or fatty acid oxidation over time.Such a dose can be determined according to the efficacy of theparticular candidate compound employed and the condition of the animal,as well as the body weight or surface area of the animal. The size ofthe dose also will be determined by the existence, nature, and extent ofany adverse side-effects that accompany the administration of acandidate compound; the LD₅₀ of the candidate compound; and theside-effects of the candidate compound at various concentrations. Ingeneral, the dose will range from 0.1-50 mg per kg, preferably 1-25 mgper kg, most preferably 1-20 mg per kg body weight. The determination ofdose response relationships is well known to one of ordinary skill inthe art.

[0189] Body Fat Reduction

[0190] Body weight reduction is typically determined by directmeasurements of the change in body fat or by loss of body weight. Bodyfat and body weight of the animals is determined before, during, andafter the administration of the candidate compound. Changes in body fatare measured by any means known in the art such as, for example, fatfold measurements with calipers, bioelectrical impedance, hydrostaticweighing, or dual x-ray absorbiometry. Preferably animals demonstrate atleast 2%, 5%, 8%, or 10% loss of body fat. Changes in body weight can bemeasured by any means known in the art such as, for example, on aportable scale, on a digital scale, on a balance scale, on a floorscale, or a table scale. Preferably animals demonstrate at least 2%, 5%,10%, or 15% loss of body weight. Body weight reduction is measuredbefore administration of the candidate compound and at regular intervalsduring and after treatment. Preferably, body weight is measured every 5days, more preferably every 4 days, even more preferably every 3 days,yet more preferably every 2 days, most preferably every day.

[0191] Changes in Fatty Acid Metabolism

[0192] Changes in fatty acid metabolism can be measured, for instance,by looking at fatty acid oxidation in cells from major fat burningtissues such as, for example, liver (Beynen, et al. Diabetes 28:828(1979)), muscle (Chiasson Lab. Anat. of Rat, (1980)), heart (Flink, etal. J. Biol. Chem. 267: 9917 (1992)), and adipocytes (Rodbell J. Biol.Chem. 239: 375 (1964)), Cells may be from primary cultures or from celllines. Cells may be prepared for primary cultures by any means known inthe art including, for example, enzymatic digestion and dissection.Suitable cell lines are known to those in the art. Suitable hepatocytelines are, for example, Fao, MH1C1, H-4-II-E, H4TG, H4-II-E-C3,McA-RH7777, McA-RH8994, N1-Si Fudr, N1-Si, ARL-6, Hepa 1-6, Hepa-1c1c7,BpRcl, tao BpRcl, NCTC clone 1469, PLC/PRF/5, Hep 3B2.1-7 [Hep 3B], HepG2 [HepG2], SK-HEP-1, WCH-17. Suitable skeletal muscle cell lines are,for example, L6, L8, C8, NOR-10, BLO-11, BC3H1, G-7, G-8, C2C12, P19,So18, SJRH30 [RMS 13], QM7. Suitable cardiac cell lines are, forexample, H9c2(2-1), P19, CCD-32Lu, CCD-32Sk, Girardi, FBHE. Suitableadipocyte lines are, for example, NCTC clone 929 [derivative of StrainL; L-929; L cell], NCTC 2071, L-M, L-M(TK-) [LMTK-; LM(tk-)], A9 (APRTand HPRT negative derivative of Strain L), NCTC clone 2472, NCTC clone2555, 3T3-L1, J26, J27-neo, J27-B7, MTKP 97-12 pMp97b [TKMp97-12],L-NGC-5HT2, Ltk-11, L-alpha-1b, L-alpha-2A, L-alpha-2C, B82.

[0193] The rate of fatty acid oxidation may be measured by ¹⁴C-oleateoxidation to ketone bodies (Guzmán and Geelen Biochem. J. 287:487(1982)) and/or ¹⁴C-oleate oxidation to CO₂ (Fruebis PNAS 98:2005 (2001);Blazquez et al. J. Neurochem 71: 1597 (1998)). Lypolysis may be measuredby fatty acid or glycerol release by using appropriate labeledprecursors or spectrophotometric assays (Serradeil-Le Gal FEBS Lett 475:150 (2000)). For analysis of ¹⁴C-oleate oxidation to ketone bodies,freshly isolated cells or cultured cell lines can be incubated with¹⁴C-oleic acid for an appropriate time, such as, for example, 30, 60,90, 120, or 180 minutes. The amount of ¹⁴C radioactivity in theincubation medium can be measured to determine their rate of oleateoxidation. Oleate oxidation can be expressed as nmol oleate produced inx minutes per g cells. For analysis of lypolysis/glycerol release,freshly isolated cells or cultured cells lines can be washed thenincubated for an appropriate time. The amount of glycerol released intothe incubation media can provide an index for lypolysis.

EXAMPLES

[0194] The following examples are provided by way of illustration onlyand not by way of limitation. Those of skill will readily recognize avariety of non-critical parameters which could be changed or modified toyield essentially similar results.

Example 1 Synthesis of Fatty Acid Ethanolamide Compounds, Homologues andAnalogs

[0195] Methods for the formation of fatty acid ethanolamines fromethanolamines and the corresponding fatty acyl are relatively straightforward and known to one of ordinary skill in the art. For example,fatty acid ethanolamides may be synthesized by reacting a fatty acid orfatty acid chloride with an aminoalcohol as described by Abadjj et al.(Abadji, V., Lin, S. Y., Taha, G., Griffin, G., Stevenson, L. A.,Pertwee, R. G. & Makriyannis, A. J. Med. Chem. 37, 1889-1893 (1994)).Fatty acids may be prepared similarly to the procedure of Serdarevichand Carroll (Serdarevich, B. & Carroll, K. K. J. Lipid Res. 7, 277-284(1966)). Radioactively labeled fatty acid ethanolamides can be preparedby reaction with acyl chlorides (Nu-Check Prep, Elysian, Minn.) with[³H]ethanolamine (10-30 Ci/mmol; American Radiolabeled Chemicals, St.Louis) as described by Desarnaud, F., Cadas, H. & Piomelli, D. (1995) J.Biol. Chem. 270, 6030-6035. Compounds can be purified by flash columnchromatography or HPLC. Compound identity can be established by use ofNMR and/or gas chromatography-mass spectrometry and thin layerchromatography.

[0196] Starting reagents and materials may be purchased from AvantiPolar Lipids, Cayman Chemicals (Ann Arbor, Mich.), Nu-Check Prep,Research Biochemicals, or Sigma. Briefly, according to methods taught byGiuffrida, A. et al. (see Giuffrida, A and Piomelli, D. in Lipid SecondMessengers (Laycock, S. G. and Rubin, R. P. Eds. pp. 113-133 CRC PressLLC, Boca Raton, Fla.) and Devane et al. (Devane W., Hanus, L. et al.Science 258, 1946-1949 (1992)), unlabeled or labeled fatty acylethanolamines can be synthesized by the reaction of the correspondingfatty acyl chlorides with unlabeled or labeled ethanolamine. The fattyacid chorides can be dissolved in dichloromethane (10 mg/ml) and reactedwith ethanolamine at −0.4° C. for 15 minutes. The reaction can be quenchby the addition of purified water. After vigorous stirring the phasesare allowed to separate. The upper aqueous phase is discarded. Theorganic phase is washed twice with water. These washes remove theunreacted ethanolamine. This method provides a quantitative formation offatty acyl ethanolamines. The ethanolamines are concentrated to drynessunder a stream of nitrogen gas and can be reconstituted in an organicsolvent such as dichloromethane at a concentration of 20 mM. Theresulting fatty acyl ethanolamine solution can be stored at −20° C.until needed for use.

[0197] The chemistry of fatty acid carboxylic acid groups, primary andsecondary amines, and primary alcohol groups is well known to one ofordinary skill in the art. Fatty acid ethanolamides having a variety ofsubstituents on the ethanolamine portion thereof can be formed in manyways, but most preferably by starting with the corresponding substitutedethanolamine and fatty acid moieties. Such substituted ethanolamineswould include the alkyl aminoethanol ethers and acyl aminoethanol estersas well as secondary akyl ethanol amines. Alternatively, the particularfatty acid ethanolamide can be synthesized from the corresponding fattyacid ethanolamide by the addition of the appropriate substituent groups.

[0198] Example 2

Methods for Screening Fatty Acid Ethanolamide (FAE) in vivo and OtherCompounds of the Invention.

[0199] Animals. Male Wistar rats (200-350 g) were used. Proceduresshould met NIH guidelines detailed in the Guide for the Care and Use ofLaboratory Animals, and the European Communities directive 86/609/EECregulating animal research.

[0200] Chemicals. FAEs and [²H₄] FAEs were synthesized in the laboratory(Giuffrida et al., “Lipid Second Messengers” (ed. Laychock, S. G. &Rubin, R. P.) 113-133 (CRC Press LLC, Boca Raton, Fla., 1998));1,2-dioleyl-sn-glycero-phosphoethanolamine-N-oleyl was purchased fromAvanti Polar Lipids (Alabaster, Ala.); SR141716A was provided by RBI(Natick, Mass.) as part of the Chemical Synthesis Program of the NIMH(N01MH30003); SR144528 was a generous gift of Sanofi Recherche; allother drugs were from Tocris (Ballwin, Mo.) or Sigma (Saint Louis, Mo.).FAE were dissolved in dimethylsulphoxide (DMSO) and administered in 70%DMSO in sterile saline (acute treatments) or 5% Tween 80/5%propylenglycol in sterile saline (subchronic treatments) (1 ml per kg,i.p.). Capsaicin was administered in 10% Tween 80/10% ethanol/80%saline; SR141716A, SR144528, CCK-8 and CP-93129 in 5% Tween 80/5%propylenglycol/90% saline (1 ml per kg, i.p.).

[0201] Enzyme assays. In all biochemical experiments, rats were killedand tissues collected between 1400 and 1600 h, after varying periods offood deprivation. Microsome fractions were prepared as described(Désarnaud et al., J. Biol. Chem., 270:6030-6035 (1995)). NAT assayswere performed using 1,2-di[¹⁴C]palmityl-sn-glycerophosphocholine as asubstrate (108 mCi/mmol, Amersham, Piscataway, N.J.) (Cadas et al., H.,J. Neurosci., 17:1226-1242 (1997)). FAAH assays were performed accordingto (Désarnaud et al., J. Biol. Chem., 270:6030-6035 (1995)), except that[³H]anandamide (arachidonyl-[1-³H]ethanolamide; 60 Ci/mmol; ARC, St.Louis, Mo.) was included as a substrate and radioactivity was measuredin the aqueous phase after chloroform extraction.

[0202] HPLC/MS analyses. Plasma was prepared from blood obtained bycardiac puncture (Giuffrida et al., Anal. Biochem., 280:87-93 (2000))and CSF was collected from the cisterna magna using a 27G 1/2 needle(Precisionglide, USA). FAEs and NAPE were extracted from tissues withmethanol/chloroform and fractionated by column chromatography (Giuffridaet al., “Lipid Second Messengers” (ed. Laychock, S. G. & Rubin, R. P.)113-133 (CRC Press LLC, Boca Raton, Fla., 1998)). FAEs were quantifiedby HPLC/MS, using an isotope dilution method (Giuffrida et al., Anal.Biochem., 280:87-93 (2000)). Individual NAPE species were identified andquantified by HPLC/MS, using an external standard method (Calignano etal., Nature, 408:96-101 (2000)).

[0203] Blood chemistry. Plasma β-hydroxybutyrate and glycerol weremeasured using commercial kits (Sigrna, St. Louis, Mo.). Plasmaprolactin, corticosterone and luteinizing hormone were quantified byradioimmunoassay (Navarro et al., Neuroreport, 8:491-496 (1997)).

[0204] Feeding experiments. Acute experiments. Food intake was measuredin 24-h food-deprived rats (Navarro et al., J. Neurochem., 67:1982-1991(1996)), administering drugs 15 min before food presentation. Subchronicexperiments. Ad libitum fed rats received vehicle injections for threedays. On day four, the animals were divided in two equal groups and gavethem daily injections of vehicle or OEA (5 mg per kg at 1900 h) for 7consecutive days, while measuring body weight, food intake and waterintake.

[0205] Conditioned taste aversion. Rats were water-deprived for 24 h andthen accustomed to drink from a graded bottle during a 30-min testperiod for four days. On day five, water was substituted with a 0.1%saccharin solution and, 30 min later, the animals received injections ofvehicle, OEA (20 mg per kg) or lithium chloride (0.4 M, 7.5 ml per kg).During the following two days, water consumption was recorded over30-min test periods. The animals were then presented with water orsaccharin, and drinking measured.

[0206] Operant responses for food. Rats were trained to lever press forfood on a fixed ratio 1 (FR1) schedule of reinforcement, whilefood-restricted at 20 g of chow per rat per day (Rodriguez de Fonseca etal., Acta Pharmacol. Sin., 20:1109-1114 (1999)). Once stable respondingwas achieved, the animals were trained to acquire an FR5, time out 2-minschedule of food reinforcement and kept in limited access to food. Whena stable baseline was obtained, the animals were used to test theeffects of vehicle or OEA (1, 5 or 20 mg per kg) administered 15 minbefore lever presentation. Test duration was 60 min.

[0207] Other behavioral assays. The elevated plus maze test wasconducted as described (Navarro et al., Neuroreport, 8:491-496 (1997))after the administration of vehicle or OEA (20 mg per kg, i.p.).Horizontal activity in an open field (Beltramo et al., J. Neurosci.,20:3401-3407 (2000)) and pain threshold in the hot plate test (55° C.)(Beltramo et al., Science, 277:1094-1097 (1997)) were measured 15 minafter injection of vehicle or OEA (20 mg per kg). Rectal temperature wasmeasured using a digital thermometer (Martin-Calderón et al., Eur. J.Pharmacol., 344:77-86. (1998)).

[0208] In situ hybridization. Rats were accustomed to the handling andinjection procedure for five days. On day six, vehicle or drug OEA (10mg per kg, i.p.), or oleic acid (10 mg per kg) was administered, and therats killed 60 min later by decapitation under anesthesia. In situhybridization analyses were conducted using ³⁵S-labeled cRNA probes forc-fos (Guthrie et al., Proc. Natl. Acad. Sci. U.S.A., 90:3329-3333(1993)) and choline acetyl transferase (ChAT) (Lauterbom et al., BrainRes. Mol. Brain Res., 17:59-69 (1993)). Average hybridization densitieswere determined from at least three tissue sections per rat. Statisticalsignificance was evaluated using one-way analysis of variance (ANOVA)followed by the Tukey-Kramer post-hoc test for paired comparisons.

[0209] Data analysis. Results are expressed as mean ±s.e.m of n separateexperiments. The significance of differences among groups was evaluatedusing ANOVA followed by a Student-Newman-Keuls post hoc test, unlessindicated otherwise.

Example 3 Effects of Starvation on OEA and Other FAE Levels in the Rat

[0210] In one embodiment, the invention provides methods of treatmentwherein individuals needing to lose weight and/or body fat are testedfor OEA levels before and/or during fasting. Individuals with low levelsof OEA prior to or in response to fasting are particularly then targetedfor OEA treatment.

[0211] Rats were deprived of food while periodically measuring FAElevels in cardiac blood by high-performance liquid chromatography (HPLC)coupled to electrospray mass spectrometry (MS). Plasma OEA remained atbaseline levels for the first 12 h of fasting, markedly increased at18-24 h, and returned to normal at 30 h (FIG. 1a). No such effect wasobserved following water deprivation (FIG. 1b) or application ofstressors such as restraint immobilization and lipopolysaccharide (LPS)administration [in pmol per ml; 10.3±0.8; 60 min after a 15-minimmobilization, 8.4±1.6; 60 min after LPS injection (1 mg per kg),7.0±0.7; n=6-9]. Plasma PEA was not significantly affected by any ofthese treatments (data not shown), whereas anandamide decreased rapidlyupon food removal, remaining lower than baseline for the entire durationof the experiment (FIG. 1d). Anandamide levels also declined afterimmobilization (in pmol per ml; control, 3.6±0.4; immobilization,1.1±0.5; n=7-8; P<0.01), LPS treatment (control, 2.0±0.5; LPS, 0.2±0.2;n=6; P<0.01) and, though not significantly, water deprivation (FIG. 1e).These results indicate that circulating OEA levels increase transientlyduring starvation. This response is selective for OEA over anandamideand other FAEs, and coincides temporally with the rise in blood glyceroland β-hydroxybutyrate (Table 1), which signals the shift of energymetabolism from carbohydrates to fatty acids as primary fuel (Cahill, G.F., Clin. Endocrinol. Metab., 5:397-415 (1976)). TABLE 1 Plasma level ofβ-hydroxybutyrate (β-HBA) and glycerol in fasting rats. β-HBA GlycerolFree feeding 1.2 ± 0.4 4.6 ± 0.9  2 h fasted 1.2 ± 0.2 5.3 ± 0.6  4 hfasted 0.8 ± 0.1 9.1 ± 1.8  8 h fasted 1.3 ± 0.2 6.3 ± 0.4 12 h fasted 4.6 ± 0.8* 7.6 ± 1.0 18 h fasted  6.8 ± 0.4*  8.4 ± 0.4* 24 h fasted 9.1 ± 1.2*  8.4 ± 0.3*

[0212] OEA levels in cerebrospinal fluid were not significantly affectedby food deprivation (FIG. 1c), implying that the surge in plasma OEA mayoriginate outside the CNS. To test this hypothesis, the impact ofstarvation on OEA metabolism in various rat tissues was investigated.The biochemical route by which animal cells produce and degrade OEA andother FAEs is thought to comprise three key enzymatic steps. Calciumion-stimulated NAT activity transfers a fatty acid group from the sn-1position of a donor phospholipid to the primary amine ofphosphatidylethanolamine, producing NAPE2 (Schmid et al., Chem. Phys.Lipids, 80:133-142 (1996); Piomelli et al., Neurobiol. Dis., 5:462-473(1998)). Cleavage of the distal phosphodiester bond in NAPE by anunknown phospholipase D generates FAEs (Schmid et al., Chem. Phys.Lipids, 80:133-142 (1996); Piomelli et al., Neurobiol. Dis., 5:462-473(1998)), which are eventually broken down to fatty acid and ethanolamineby an intracellular fatty acid amide hydrolase (FAAH) (Schmid et al., J.Biol. Chem., 260:14145-14149 (1985); Cravatt et al., Nature, 384:83-87(1996)). Food deprivation (18 h) was accompanied by a marked increase inNAT activity in white adipose tissue (FIG. 2a), but not in the brain,stomach or kidney (FIGS. 2b,d and data not shown). In liver, intestinesand skeletal muscle, NAT activity was reduced by fast (FIGS. 2c,d anddata not shown). These enzymatic changes were paralleled bycorresponding alterations in NAPE tissue content. Several molecularspecies of NAPE are present in rat tissues, including the OEA precursorsalk-1-palmitoenyl-2-arachidonyl-sn-glycero-phosphoethanolamine-N-oleyl(NAPE 1; FIG. 3a) andalk-1-palmityl-2-arachidonyl-sn-glycero-phosphoethanolamine-N-oleyl(NAPE 2; FIG. 3a); and the PEA precursoralk-1-palmityl-2-arachidonyl-sn-glycero-phosphoethanolamine-N-palmityl(not shown). In agreement with NAT activity measurements, fooddeprivation increased NAPE content in fat, and decreased it in liver(FIGS. 3b,c).

[0213] Since NAPE biosynthesis and FAE formation are tightly coupledprocesses (Cadas et al., H., J. Neurosci., 17:1226-1242 (1997)), onemight expect starvation to augment the levels of OEA and other FAEs inadipose, but not in other tissues. Accordingly, fat from starved ratscontained more OEA and PEA than did fat from free-feeding controls (FIG.3d and data not shown), whereas no such difference was seen in thebrain, stomach, and intestines (data not shown). Contrary to ourexpectation, however, the liver content of OEA and PEA was also higherin food-deprived than in free-feeding rats (FIG. 3d and data not shown).This discordance may be due to an accumulation of FAEs by the liver,which is consistent with the postulated roles of this organ in FAErecapture and metabolism (Bachur et al., J. Biol. Chem., 240:1019-1024(1965); Schmid et al., J. Biol. Chem., 260:14145-14149 (1985)).

[0214] The hydrolysis to fatty acid and ethanolamine, catalyzed by FAAH,is a key step in FAE degradation (Bachur et al., J. Biol. Chem.,240:1019-1024 (1965); Schmid et al., J. Biol. Chem., 260:14145-14149(1985); Cravatt et al., Nature, 384:83-87 (1996); Désarnaud et al., J.Biol. Chem., 270:6030-6035 (1995)). Food deprivation profoundly reducedFAAH activity in adipose membranes, but had no effect on FAAH activityin the brain, liver, stomach, intestines, kidney and skeletal muscle(FIGS. 2a-e and data not shown). Thus, food deprivation may increase thelevels of OEA and other FAEs in white fat in two synergistic ways, whichare mechanistically distinct from other reactions occurring duringlipolysis: stimulation of NAT activity may lead to increase thebiosynthesis of NAPE and FAEs, while inhibition of FAAH activity mayprolong the life span of newly synthesized FAEs. Although severaltissues may contribute to the normal levels of OEA in the bloodstream,the dynamic biochemical changes observed in fat underscore the crucialrole of this tissue in generating OEA during starvation.

Example 4 Suppression of Food Intake by OEA and Other FAEs

[0215] The effects of systemically administered OEA on food intake inrats can be assessed using a 24 h fast. In this system, OEA caused adose- and time-dependent suppression of food intake (FIGS. 4a,b). Todefine the selectivity of this response, various OEA analogs wereevaluated for their ability to produce hypophagia.

[0216] Anandamide and oleic acid had no effect.

[0217] Palmitylethanolamide was active but significantly less potentthan OEA.

[0218] Elaidylethanolamide (an unnatural OEA analog) was similar inpotency to OEA (FIG. 4a).

[0219] These results indicate that OEA reduces eating in a structurallyselective manner and that other fatty acid ethanolamide-like compoundscan be identified for use according to the invention.

Example 5 Specificity Over Cannabinoid Receptor Activators

[0220] The molecular requisites for OEA hypophagia are distinct fromthose involved in the interaction of anandamide with its knowncannabinoid targets (Khanolkar et al., Life Sci., 65:607-616 (1999)).Cannabinoid receptor antagonists did not affect OEA hypophagia in vivo,and OEA did not displace cannabinoid binding to rat brain membranes invitro. Thus, despite its structural and biogenetic relationships withanandamide, OEA does not depend on the endogenous cannabinoid system toproduce anorexia.

Example 6 Sustained Body Weight Reduction

[0221] In some embodiments, the compounds of the instant inventionprovide for a sustained fat reduction or body weight reduction uponprolonged administration to mammals. This effect is advantageous as avariety of drugs suppress eating after acute administration, but fail todo so when treatment is prolonged (Blundell, J., Trends Pharmacol. Sci.,12:147-157 (1991)).

[0222] OEA was subchronically administered to rats. Daily injections ofOEA (5 mg per kg, i.p.) for seven days resulted in a small, butsignificant decrease in cumulative food intake (FIG. 5a), which wasaccompanied by a profound inhibition of weight gain (FIGS. 5b,c). OEAdid not affect water intake (FIG. 5d). The impact of OEA on body weightis only partially explained by its moderate reduction of foodconsumption indicating that other factors, such as stimulation of energyexpenditure or inhibition of energy accumulation, may contribute to thiseffect.

Example 7 FAE's May Have a Peripheral Site of Action

[0223] In one of its aspects, the invention provides compounds with aperipheral site of action. Such a site is advantageous in reducing thelikelihood of central nervous system side effects.

[0224] Though potent when administered peripherally, OEA was ineffectiveafter direct injection into the brain ventricles (Table 2), suggestingthat the primary sites of action of this compound might be locatedoutside the CNS. As a further demonstration, sensory fibers in the vagusand other peripheral nerves were chemically destroyed by treating adultrats with the neurotoxin, capsaicin (Kaneko et al., Am. J. Physiol.,275:G1056-G1062 (1998)). Capsaicin-treated rats failed to respond toperipherally administered cholecystokinin-8 (CCK-8) (FIGS. 6,a,c), drankmore water than controls (FIGS. 6b,d) and lost the corneal chemosensoryreflex (data not shown), three indications that the neurotoxin haddestroyed sensory afferents (MacLean, D. B., Regul. Pept., 11:321-333(1985); Ritter et al., Am. J. Physiol., 248:R501-R504 (1985); Curtis etal., Am. J. Physiol., 272:R704-R709 (1997)). Treated animals also failedto respond to OEA (10 mg per kg, i.p.), but responded normally to thecompound CP-93129, which targets 5-HT1B receptors in the CNS (FIGS.6a,c) (Lee et al., Psychopharmacology, 136:304-307 (1998)). Thesefindings support the hypothesis that OEA causes hypophagia by acting ata peripheral site, and that sensory fibers are required for this effect.TABLE 2 Effects of intracerebroventricular pranamide on food intake. 60min 120 min 240 min vehicle 5.8 ± 0.6 8.0 ± 0.5 9.5 ± 0.5 prana 0.4 μg4.8 ± 0.4 6.6 ± 0.4 8.4 ± 0.4 prana 2 μg 4.9 ± 0.4 6.6 ± 0.6 8.7 ± 0.5prana 10 μg 5.9 ± 0.2 8.1 ± 0.4 9.6 ± 0.7

[0225] The compounds of the invention may use peripheral sensory inputsto suppress appetite. Peripheral sensory inputs related to appetitesuppression recruit several CNS structures, which include the nucleus ofthe solitary tract (NST) in the brainstem and the arcuate andparaventricular (PVN) nuclei in the hypothalamus (Schwartz et al.,Nature, 404:661-671 (2000)). To identify the brain pathways engagedduring OEA-induced hypophagia, mRNA levels for the activity regulatedgene c-fos (Curran et al., Oncogene, 2:79-84 (1987)) were mapped by insitu hybridization after systemic administration of OEA, oleic acid orvehicle. When compared to controls, OEA (10 mg per kg, i.p.) evoked ahighly localized increase in c-fos mRNA levels in the PVN, supraopticnucleus (FIG. 7a) and NST (FIG. 7c). This enhancement was specific tothese areas, insofar as c-fos expression in other brain regions was notsignificantly affected by OEA treatment (FIGS. 7b,d). The finding thatOEA stimulates c-fos mRNA expression in the NST (which processes vagalsensory inputs to the CNS) and the PVN (a primary site for theorchestration of central catabolic signals) (Schwartz et al., Nature,404:661-671 (2000)), is consistent with a physiological role for thislipid as a peripheral mediator of anorexia.

[0226] It is possible that OEA reduced eating by inducing a non-specificstate of behavioral suppression. If this is the case, OEA should causeconditioned taste aversion, which can be readily provoked in rats by anumber of noxious substances (Green et al., Science, 173:749-751(1971)), including lithium chloride (FIG. 4c). However, a maximal doseof OEA (20 mg per kg, i.p.) had little effect in this assay (FIG. 4c),suggesting that the compound may not be aversive. Several additionalobservations support the behavioral specificity of OEA. OEA did notalter water intake, body temperature, pain threshold (FIGS. 4d-f), oractivity of the hypothalamus-pituitary-adrenal (HPA) axis (Table 3).Moreover, OEA did not produce anxiety-like symptoms (FIG. 4g) and,though it reduced motor activity and operant responses for food, it didso at a dose that was substantially higher than those required toproduce hypophagia (FIGS. 4h-i). This pharmacological profiledifferentiates OEA from other appetite suppressants such as amphetamineand glucagon-like peptide 1 (whose effects often include aversion,hyperactivity, anxiety and activation of the HPA axis) and from theendogenous cannabinoid anandamide (which stimulates food intake inpartially satiated animals, increases pain threshold, decreases bodytemperature and activates the HPA axis) (Pertwee, R. G., Exp. Opin.Invest. Drugs, 9:1553-1571 (2000)). TABLE 3 Effects of OEA on plasmahormone levels. B PRL LH vehicle 212 ± 24 10.8 ± 2.7 5.3 ± 0.9 prana 20280 ± 61  8.2 ± 3.2 6.2 ± 1.5

[0227] OEA elicits hypophagia at physiologically relevant doses. 1 hrafter administration of a half-maximally effective dose (5 mg per kg,i.p.), circulating OEA levels (16.1±2.6 pmol per ml) were significantlyhigher than baseline (10.1±1.1; P<0.05, Student's t test; n=5), butbelow those measured in 18-h food-deprived animals (FIG. 1a). Thus, theconcentrations reached by OEA in blood during starvation can besufficient to elicit notable behavioral responses.

Example 8 Identifying Body Fat Reducing Compounds of the Invention

[0228] The following example demonstrates how to identify appetitesuppressors using OEA as a positive control. In particular, thesynthesis of OEA, the measurement of body fat reduction and fatty acidoxidation are discussed. Synthesis of OEA.

[0229] Oleylchloride is purchased from Nu-Check Prep (Elysian, Minn.) orprepared following standard procedures. Oleylchloride is dissolved indichloromethane (10 mg/ml) and allowed to react with five equivalents ofethanolamine for 15 min. at 0-4° C. The reaction is stopped by theaddition of purified water. After vigorous stirring and phaseseparation, the upper aqueous phase is discarded and the organic phaseis washed twice with water to remove non-reacted ethanolamine. Theresulting OEA is concentrated to dryness under a N₂ stream,reconstituted in chloroform at 20 mM, and stored at −20° C. until use.

[0230] Measuring Body Fat Reduction Induced by Candidate Compounds

[0231] The ability of a compound to reduce body fat can be evaluated bya number of methods. For example, appropriate amounts OEA and/orcandidate compounds are administered to rats via intraperitonealinjection. The OEA and candidate compounds can be formulated in 70% DMSOin sterile saline, 5% Tween 80/5% propylenglycol in sterile saline, or10% Tween 80/10% ethanol/80% saline. Five mg per kg of OEA can be usedas the positive control. Amounts of candidate compounds administered mayrange, for instance, from 1-25 mg per kg. Typically 1, 2, 5, 10, 15, and20 mg per kg doses of each candidate compound can be administered todifferent sets of rats to determine which dose is optimal. Injectionsmay be given 30 minutes before the animals' principal meal for 7-14days.

[0232] The effect of the candidate compound on total body fat can bedetermined by taking direct measurements of the rat's body fat usingskin fold calipers. Skin on the rats' backs, abdomen, chest, front andrear legs can be pinched with calipers to obtain measurements beforeadministration of OEA and/or candidate compounds and every 48 hoursduring and after administration of OEA and/or candidate compounds.Differences in measurements in at least two of the pinched sites reflectthe change in the rat's total body fat.

[0233] Measuring Fatty Acid Oxidation Induced by Candidate Compounds

[0234] Compounds can also be assayed for their effect on fatty acidmetabolism. The effect of the candidate compound on fatty acidmetabolism can be measured by measurements of fatty acid oxidation inprimary cultures of liver cells. Hepatocytes may be used to determinethe rate of oleate oxidation to ketone bodies and carbon dioxide. Suchcells can be isolated from adult rat liver by enzymatic digestion asdescribed by Beynen et al. in Diabetes 28:828 (1979). Cells typicallyare cultured in suspension and incubated in Krebs-Henseleit'sbicarbonate medium supplemented with bovine serum albumin and glucose asdescribed by Guzmán & Geelen, Biochem. J. 287:487(1992). The proteinconcentration of the cultured cells can be determined and cells seededin 2 ml media so that 4-6 mg protein per ml is present in the reactionmixture. Cells can be incubated for 10 minutes at 37° C. with[¹⁴C]-oleic acid (Arnersham), in the presence or absence of 10 μM OEA,reactions may be stopped with 200 μl 2M perchloric acid and acid-solubleproducts extracted with chloroform/methanol/water (5:1:1, vol:vol:vol).The aqueous phase can be removed and washed twice more. Proteinconcentration can be determined using a Lowry assay. The rate of oleateconversion into ketone bodies may be expressed as rnmol of oleateoxidized per hour per mg protein and may be determined using liquidscintillation counting. Accordingly, OEA enhances oleate oxidation by21+−6% (n=4, p<0.01 vs. control incubations by the Student t test).

Example 9 Effect of OEA on Fatty Acid Metabolism

[0235] Oleoylethanolamide (OEA) decreases body weight not only bysuppressing appetite, but also by possibly enhancing body fatcatabolism. The effects of OEA on fatty acid oxidation in major body-fatburning tissues (soleus muscle, liver, cultured cardiac myocytes andastrocytes) was examined. OEA significantly stimulates fatty acidoxidation in primary cultures of liver, skeletal muscle (soleus) andheart cells, whereas it has no effect in brain-derived astroglial cellcultures. In addition, OEA induces a significant mobilization oftriacylglycerol stores from primary white adipose tissue cells. Table 4details the methods and effects of OEA on fatty acid oxidation in thesecells. Structure-activity relationship experiments provide evidence thatthe effect of OEA on skeletal muscle fatty acid oxidation is specific(FIG. 8). Thus, the effects of OEA are mimicked by thehydrolysis-resistant homologue methyl-OEA and -only partially- bypalmitylethanolamide (PEA), but not by arachidonylethanolamide (AEA) oroleic acid (OA). In short, these results show that lipid oxidation andmobilization are enhanced by OEA, and that the effects of OEA arerestricted to peripheral sites. TABLE 4 Cell/tissue Hepatocyte Soleusmuscle Cardiomyocyte Astrocyte Adipocyte Origin Adult rat Adult ratNewborn rat Newborn rat Adult rat liver hind limb heart brain cortexepididymus Isolation Enzymatic Dissection Enzymatic Enzymatic Enzymaticprocedure digestion (Chiasson, digestion digestion digestion (Beynen et1980) (Flink et (McCarthy & (Rodbell, al., 1979) al., 1992) De Vellis,1964) 1980) Type of Cell Tissue Cell Cell Cell culture suspensionsuspension monolayer monolayer suspension Incubation Krebs- Krebs-Hen-High-glucose Hams Krebs- medium Henseleit seleit Hepes DMEM plusF12/DMEM Henseleit bicarbonate plus BSA BSA plus insulin, Hepes plusplus BSA and glucose (Wu et al., transferrin, BSA and and glucose(Fruebis 2000) progesterone, glucose (Guzman & et al., putrescine(Rodbell, Geelen, 1992) 2001) and selenite 1965) (Blazquez et al., 1998Metabolic [¹⁴C]oleate [¹⁴C]oleate [¹⁴C]oleate [¹⁴C]oleate Lypolysisparameter oxidation to oxidation to oxidation to oxidation to (glycerolketone bodies CO₂ (Fruebis et CO₂ (Blazquez ketone release) (Guzman &al., 2001) et al., 1998) bodies (Blaz- (Serradeil- Geelen, 1992) quez etal., Le Gal et 1998) al., 2000) Incubation 10 30 30 30 30 time (min)Stimulatory 21 ± 6 36 ± 10 37 ± 9 2 ± 6 38 ± 16 effect of 10 (n = 4) (n= 4) (n = 3) (n = 3) (n = 3) μM OEA (%) Statistical P < 0.01 P < 0.01 P< 0.01 Non P < 0.01 significance significant vs. control #Rodbell M JBiol Chem 239: 375-380 (1964); Rodbell M Ann NY Acad Sci 131: 302-314(1965); Serradiel-Le Gal C et al., FEBS Left 475: 150-156 (2000); Wu Wet al., J Biol Chem 275: 40133-40119 (2000).

Example 10 Role of Endogenous OEA in the Intestines

[0236] The impact of feeding on intestinal OEA biosynthesis was studied.High performance liquid chromatography/mass spectrometry analysesrevealed that small intestinal tissue from free-feeding rats containssubstantial amounts of OEA (354±86 pmol per g, n=3). Intestinal OEAlevels were markedly decreased after food deprivation, but returned tobaseline after refeeding. By contrast, no such changes were observed instomach (in pmol per g; control, 210±20; starvation, 238±84;starvation/refeeding, 239±60, n=3). Variations in intestinal OEA levelswere accompanied by parallel alterations in NAT activity, whichparticipates in OEA formation, but not in fatty acid amide hydrolaseactivity, which catalyzes OEA hydrolysis. These findings suggest thatstarvation and feeding reciprocally regulate OEA biosynthesis in smallintestine. In agreement with an intra-abdominal source of OEA, plasmaOEA levels in starved rats were found to be higher in portal than incaval blood (in pmol per ml; porta, 14.6±1.8; cava, 10.3±2.8; n=5). Thecontribution of other intra-abdominal tissues to OEA formation cannot beexcluded at present. These results suggest many interventions to utilizethe OEA systems in feeding behavior. According to this model, foodintake may stimulate NAT activity enhancing OEA biosynthesis in thesmall intestine and possibly other intra-abdominal tissues. Newlyproduced OEA may activate local, sensory fibers, which may in turninhibit feeding by engaging brain structures such as the NST and PVN.

[0237] Our results reveal an unexpected role for OEA in the peripheralregulation of feeding, and provide a framework to develop novelmedicines for reducing body weight or body fat, for preventing bodyweight gain or body fat increase, for suppressing appetite or reducingfood seeking behavior, or food intake, and for the treating eatingdisorders, overweight, or obesity. These medicines would include notonly OEA analogues and homologues but also agents which controlling OEAlevels by acting upon the OEA formation and hydrolyzing systems andenzymes as disclosed above.

[0238] All publications and patent applications cited in thisspecification are herein incorporated by reference to the extent notinconsistent with the present disclosure as if each individualpublication or patent application were specifically and individuallyindicated to be incorporated by reference.

[0239] Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it will be readily apparent to those of ordinary skill inthe art in light of the teachings of this invention that certain changesand modifications may be made thereto without departing from the spiritor scope of the appended claims.

What is claimed is:
 1. A method of reducing food intake in a mammal,said method comprising administering to said mammal a fatty acidalkanolamide, wherein said administering is in an effective amount toreduce food intake in said mammal.
 2. The method according to claim 1,wherein the fatty acid alkanolamide is oleoylethanolamide.
 3. The methodof claim 1, wherein the fatty acid alkanolamide comprises a fatty acidmoiety covalently bonded to an ethanolamine moiety via an amide linkage.4. The method of claim 3, wherein the fatty acid moiety ismonounsaturated or polyunsaturated.
 5. The method of claim 1, whereinthe administering is via a dermal patch.
 6. The method of claim 4,wherein the fatty acid moiety is oleic acid.
 7. The method of claim 3,wherein the fatty acid moiety has from 12 to 20 carbon atoms.
 8. Themethod of claim 3, wherein the fatty acid is selected from the groupconsisting of elaidic acid, palmitoleic acid, palmitic acid, linoleicacid, alpha-linolenic acid, and gamma-linolenic acid.
 9. The method ofclaim 3, wherein the hydroxy group of the ethanolamine moiety issubstituted with a lower (C₁-C₃) alkyl group to form the correspondingether.
 10. The method of claim 3, wherein the hydroxy group of theethanolamine moiety is bound to a the carboxylate group of a lower(C₂-C₆) alkyl carboxylic acid to form the corresponding ester.
 11. Themethod of claim 3, wherein the fatty acid ethanolamide further comprisesa lower alkyl (C₁-C₃) group covalently bound to the nitrogen atom of thefatty acid ethanolamide.
 12. A method according to claim 1, wherein themammal is human.
 13. A method according to claim 1, wherein the fattyacid alkanolamide is palmitoylethanolamide.
 14. A method according toclaim 1, wherein the fatty acid alkanolamide does not activate thecannabinoid CB2 or the cannabinoid CB1 receptor.
 15. A method accordingto claim 1, wherein said fatty acid alkanolamide is administered with apharmaceutically acceptable carrier by an oral, rectal, topical, orparenteral route.
 16. A method of reducing or controlling body fat orbody weight in a mammal, said method comprising administering to saidmammal, in an effective amount to reduce body fat or body weight, acompound of the formula

or a pharmaceutically acceptable salt thereof, wherein n is from 0 to 5,the sum of a and b can be from 0 to 4; Z is a member selected from thegroup consisting of —C(O)N(R^(o))—; —(R^(o))NC(O)—; —OC(O)—; —(O)CO—; O;NR^(o); and S; and wherein R^(o) and R² are members independentlyselected from the group consisting of unsubstituted or unsubstitutedalkyl, hydrogen, C₁-C₆ alkyl, and lower (C₁-C₆) acyl, and wherein up tofour hydrogen atoms of the fatty acid portion and alkanol portionthereof are substituted by methyl or a double bond, and the bond betweencarbons c and d may be unsaturated or saturated.
 17. A method of claim16, said compound is of the formula:

or a pharmaceutically acceptable salt thereof, wherein n is from 0 to 4,the sum of a and b is from 1 to 3, and R¹ and R² are membersindependently selected from the group comprising hydrogen, C₁-C₆ alkyl,and lower (C₁-C₆) acyl, and wherein up to four hydrogen atoms of thefatty acid portion and alkanolamine portion thereof are substituted bymethyl or a double bond, and the bond between carbons c and d may beunsaturated or saturated.
 18. The method of claim 17, wherein R¹ and R²are members independently selected from the group comprising hydrogen,C₁-C₃ alkyl, and lower (C₁-C₃) acyl.
 19. The method of claim 17, whereina=1 and b=1.
 20. The method of claim 17, wherein n=1.
 21. The method ofclaim 17, wherein R¹ and R² are each H.
 22. The method of claim 17,wherein the bond between carbon c and carbon d is a double bond.
 23. Themethod of claim 17, wherein the compound is oleoylethanolamide.
 24. Themethod of claim 17, wherein the compound is palmitoylethanolamide 25.The method of claim 17, wherein the administering is parenteral, oral,transdermal, rectal, or intranasal.
 26. The method of claim 17, whereinthe mammal is a human.
 27. The method of claim 16, wherein the compoundis according to one of the following formulae:

wherein n is from 1-5 and the sum of a and b is from 0 to 4; R² isselected from the group consisting of hydrogen, C₁-C₆ alkyl, and lower(C₁-C₆) acyl; and up to four hydrogen atoms of the fatty acid portionand alkanol portion thereof may also be substituted by methyl or adouble bond.
 28. A pharmaceutical composition comprising apharmaceutically acceptable excipient and a compound of the formula:

or a pharmaceutically acceptable salt thereof, wherein n is from 0 to 5,the sum of a and b can be from 0 to 4; Z is a member selected from thegroup consisting of —C(O)N(R)—; —(R^(o))NC(O)—; —OC(O)—; —(O)CO—; O; NR;and S; and wherein R^(o) and R² are members independently selected fromthe group consisting of unsubstituted or unsubstituted alkyl, hydrogen,C₁-C₆ alkyl, and lower (C₁-C₆) acyl, and wherein up to four hydrogenatoms of the fatty acid portion and alkanol portion thereof aresubstituted by methyl or a double bond, and the bond between carbons cand d may be unsaturated or saturated.
 29. A pharmaceutical compositionof claim 28, wherein the compound is of the formula:

or a pharmaceutically acceptable salt of the compound, wherein n is from1 to 3, the sum of a and b is from 1 to 3, and R and R² are membersindependently selected from the group consisting of hydrogen, C₁-C₆alkyl, and lower (C₁-C₆) acyl and wherein up to four hydrogen atoms ofthe fatty acid portion and alkanol portion thereof are substituted bymethyl or a double bond, and the bond between carbons c and d may beunsaturated or saturated.
 30. The composition of claim 29, wherein saidcomposition is in unit dosage format and the unit dose contains aneffective amount of the compound to reduce or 3 control body weight. 31.The composition of claim 29, wherein the amount is about from 10 to 1000mg.
 32. The composition of claim 29, wherein the amount is about from 1to 100 mg.
 33. The composition of claim 29, wherein the amount is aboutfrom 100 to 500 mg.
 34. The composition of claim 29, wherein thecompound is palmitoylethanolamide.
 35. The composition of claim 29,wherein the composition is a topical composition, an oral composition,or a parenteral composition.
 36. The composition of claim 29, wherein R¹and R² are members independently selected from the group comprisinghydrogen, C₁-C₃ alkyl, and lower (C₁-C₃) acyl.
 37. The composition ofclaim 29, wherein a=1 and b=1 and n=1.
 38. The composition of claim 29,wherein the compound is oleoylethanolamide.
 39. The composition of claim29, wherein R¹ and R² are each H.
 40. The composition of claim 30,wherein the bond between carbon c and carbon d is a double bond.
 41. Thecomposition of claim 30, wherein the compound is palmitylethanolamide.42. The pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound of the formula:

wherein the sum of a and b is from 0 to 4, and up to four hydrogen atomsof the fatty acid portion of the above formula may also be substitutedby methyl or a double bond; and the molecular bond between carbons c andd may be unsaturated or saturated and R represents group selected fromthe group consisting of straight and branched chain alkyl amines, acyclic alkyl amine, a furan, tetrahydrofuran, a pyrole, pyrollidine, anda pyridine; and wherein the compound is present in an effective amountfor reducing food intake upon administration to a mammal.
 43. Apharmaceutical composition comprising a pharmaceutically acceptableexcipient and a fatty acid alkanolamide in an effective amount forreducing body weight upon administration to a mammal.
 44. Thecomposition of claim 46, wherein the fatty acid moiety is oleic acid.45. The composition of claim 46, wherein the fatty acid moiety has from12 to 20 carbon atoms.
 46. The composition of claim 46, wherein thefatty acid is selected from the group consisting of elaidic acid,palmitoleic acid, palmitic acid, linoleic acid, alpha-linolenic acid,and gamma-linolenic acid.
 47. The composition of claim 46, wherein thealkanolamine of the alkanolamide is ethanolamine.
 48. The composition ofclaim 46, wherein the composition is an enteric-coated oral formulation.